Phospho-RB1 (Ser807) Antibody
Description
Introduction to Phospho-RB1 (Ser807) Antibody
Phospho-RB1 (Ser807) Antibody is a polyclonal rabbit IgG antibody that selectively recognizes RB1 phosphorylated at serine 807 (and often serine 811 due to sequence homology). This antibody is widely used in cancer research to study RB1 inactivation mechanisms, which are pivotal for understanding cell cycle progression and tumor suppression .
Role in Cell Cycle Regulation
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RB1 phosphorylation at Ser807/811 is linked to cell cycle progression. Cyclin-dependent kinases (CDKs) phosphorylate RB1, releasing E2F transcription factors to drive G1/S transition .
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SMYD2-mediated monomethylation of RB1 at lysine 810 enhances phosphorylation at Ser807/811, promoting cancer cell proliferation .
Apoptosis Regulation
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Phosphorylated RB1 (Ser807/811) binds Bax, a pro-apoptotic protein, to inhibit apoptosis. Dephosphorylation of RB1 dissociates this complex, triggering cell death .
Cancer Relevance
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Elevated phosphorylated RB1 levels are observed in cancers, making this antibody valuable for studying tumorigenesis and therapeutic targets .
Key Applications
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Western Blot (WB): Detects endogenous phosphorylated RB1 at 110 kDa in calyculin A-treated Jurkat cells .
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Immunohistochemistry (IHC): Validated in paraffin-embedded human breast carcinoma tissues .
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Immunoprecipitation (IP): Confirms RB1-Bax interactions in MCF7 and Hs578T cells .
Mechanistic Insights
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Cell Cycle Control: RB1 phosphorylation at Ser807/811 disrupts its binding to E2F, enabling transcription of S-phase genes .
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SMYD2 Interaction: SMYD2 methylates RB1 at lysine 810, priming Ser807/811 phosphorylation and accelerating G1/S transition .
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Apoptosis Suppression: Phosphorylated RB1 sequesters Bax, blocking apoptosis in cancer cells .
Table 2: Functional Outcomes of RB1 Phosphorylation
Product Specs
Target Background
- Concurrent mutations in genes such as CDKN2B or RB1 were associated with worse clinical outcomes in lung adenocarcinoma patients with EGFR active mutations. PMID: 29343775
- Mutational screening of the germline RB1 gene in Vietnamese patients with retinoblastoma reveals three novel mutations. PMID: 29568217
- Analyses with phospho-defective and phospho-mimetic mutants of FoxM1b identified a critical role of the Plk1 phosphorylation sites in regulating the binding of FoxM1b to Rb and DNMT3b. PMID: 28387346
- The accumulation of sequence variations in the RB1 gene might influence Greek patients' susceptibility towards the progression of cervical neoplasia. PMID: 30303478
- Vitiligo lesions exhibited dysregulated SUMOylation and deSUMOylation in keratinocytes. Dysregulation of the cell cycle progression was observed in SUMO1 knockdown HaCaT cells, suggesting that the deSUMOylation of Rb in keratinocytes may play a significant role in the development of vitiligo. PMID: 30066925
- The Rb1 tumor suppressor gene modifies telomeric chromatin architecture by regulating TERRA expression. PMID: 28169375
- These findings demonstrate that developmental stage-specific, as well as species- and cell type-specific features sensitize to RB1 inactivation, revealing the human cone precursors' capacity to model retinoblastoma initiation, proliferation, premalignant arrest, and tumor growth. PMID: 30213853
- Low pRB expression is associated with mouth cancer. PMID: 30275188
- Control of the Restriction Point by Rb and p21. PMID: 30111539
- Results showed that (a) alterations of the p53 and Rb pathways are associated with high proliferation of tumor cells in buccal squamous cell carcinoma (BUC) and (b) high expression of cell-cycle proteins is associated with adverse histopathological parameters of these tumors. PMID: 29970521
- The present result indicated that vascular smooth proliferation is regulated by activation of the NF-kappaB p65/miR17/RB pathway. As NF-kappaB p65 signaling is activated in and is a master regulator of the inflammatory response, these findings may provide a mechanism for the excessive proliferation of VSMCs under inflammation during vascular disorders and may identify novel targets for the treatment of vascular diseases. PMID: 29115381
- Reduced RB expression in medullary thyroid cancer is associated with decreased patient survival in univariate and multivariable analyses, independent from patient age at surgery or advanced TNM stage. PMID: 29105562
- According to immunohistochemistry and immunoblot analysis, the expression levels of cyclin D1, cyclin E, pRb, and Ki67 in psoriasis lesions decreased after treatment and were similar to those in the normal group. PMID: 29115643
- Data indicate that nuclear envelope rupture in cancer cells is likely due to the loss of either the Rb or the p53 pathway. PMID: 28811362
- Altered pRb is frequently expressed in gastric carcinoma and inversely correlates with tumor invasion and tumor stage, suggesting an early event in gastric carcinogenesis. PMID: 28965621
- Results define a network of E2F target genes as susceptible to the regulatory influence of H1.2, where H1.2 augments global association of pRb with chromatin, enhances transcriptional repression by pRb, and facilitates pRb-dependent cell-cycle arrest. PMID: 28614707
- The increased expression of miR-503-5p significantly reduced the expressions of E2F transcription factor 3 (E2F3) mRNA and retinoblastoma protein (Rb)/E2F signaling pathway mRNA in bladder cancer cells. PMID: 29169421
- Loss of Rb immunolabeling and KRAS mutation are promising molecular markers of the therapeutic response to platinum-based chemotherapy for pancreatic neuroendocrine neoplasm grade-3 (PanNEN-G3), and Rb for neuroendocrine tumor with G3 (NET-G3). PMID: 28455360
- We recommend intensive ocular screening for patients with germline RB1 mutations for retinoblastoma as well as neuroimaging for pineoblastoma surveillance. There is an approximately 20% risk of developing second primary cancers among individuals with hereditary RB, higher among those who received radiotherapy for their primary RB tumors. PMID: 28674118
- The SNPs rs 216311, rs 1800383, and rs 1800386 associated significantly with bleeding in study subjects. rs1800386 occurred in all with bleeding history; no ethnic variations were noted. PMID: 28091443
- miR-215 promoted cell migration and invasion of gastric cancer by directly targeting RB1. PMID: 28689850
- MiR-661 promotes metastasis of non-small cell lung cancer through RB/E2F1 signaling and epithelial-mesenchymal transition events. PMID: 28716024
- RB1 was identified as a direct and functional target of miR-215. RB1 is generally down-regulated in glioma tissues, and its expression inversely correlated with miR-215, which is up-regulated in high-grade glioma tissues, and its expression was negatively correlated with miR-215. PMID: 28573541
- Loss of retinoblastoma in pleomorphic fibroma: An immunohistochemical and genomic analysis. PMID: 28543636
- Results show that RB1 expression is regulated by cdc37, which facilitates its phosphorylation through increasing CDK4 stability. PMID: 29288563
- SOX2 overexpression and the loss of Rb1 protein expression might have a pivotal role in the divergent differentiation of pluripotent embryonic-like epithelial cells and the development of esophageal small-cell carcinoma. PMID: 28106103
- Several RB1 alterations associated with retinoblastoma in humans were present in several non-human primates without an apparent pathological effect. PMID: 28401291
- Results suggest that RB1 is the dominant tumor suppressor PP in Merkel cell carcinoma (MCC), and that inactivation of RB1 by MCPyV-LT is largely sufficient for its growth-supporting function in established MCPyV-positive MCC cells. PMID: 27121059
- The frequency and association of polymorphisms in the TP53 and RB1 genes with clinical characteristics in a group of children with retinoblastoma (RB) in northern Mexico were examined. PMID: 28210099
- RB underexpression is associated with tumor cell invasiveness and neuroendocrine differentiation in prostate cancer. PMID: 27015368
- Authors show that MYC inhibition by Omomyc, a dominant-negative MYC, suppresses the growth of small cell lung cancer (SCLC) cells with TP53 and RB1 inactivation carrying MYC, MYCL, or MYCN amplification. PMID: 27105536
- Data suggest that the platelet-derived growth factor receptor alpha (PDGFRalpha)/Stat3 transcription factor/Rb1 protein regulatory axis might represent a potential therapeutic target for glioblastoma (GBM) treatment. PMID: 27344175
- miR-590 inhibits RB1 and promotes proliferation and invasion of T-cell acute lymphoblastic leukemia cells. PMID: 27036041
- Causative RB1 mutations in most bilateral retinoblastoma (RB) patients and in some unilateral RB patients, including five novel mutations, were identified. PMID: 29261756
- Homozygous loss of RB1 is an independent prognostic marker in multiple myeloma. PMID: 28234347
- In certain contexts, Rb loss enables TRbeta1-dependent suppression of SKP2 as a safeguard against RB1-deficient tumorigenesis. TRbeta2 counteracts TRbeta1, thus disrupting this safeguard and promoting the development of RB1-deficient malignancies. PMID: 28972075
- Expression levels of miR-675-5p in glioma tissues and cells were negatively correlated with RB1 expression at both mRNA and protein levels and promoted cell proliferation and migration. PMID: 28970140
- Disruption of DREAM and RB-E2F complexes by oncoproteins from DNA tumor viruses leads to the upregulation of cell cycle genes and impairs growth-inhibiting pathways, including the p53-mediated downregulation of cell cycle genes. [review] PMID: 28799433
- A relatively stable genome in retinoblastoma tumor cells is maintained by TRb1 and TRb2-mediated PTTG1 inhibition, counteracting Rb-deficiency-related genomic instability. PMID: 28242412
- APC/C and pRB interact with each other via the co-activator of APC/C, FZR1, providing an alternative pathway of regulation of G1 to S transition by pRB using a post-translational mechanism. Both pRB and FZR1 have complex roles and are implicated not only in regulation of cell proliferation but also in differentiation, quiescence, apoptosis, maintenance of chromosomal integrity, and metabolism. PMID: 27402801
- Analysis of the spectrum of RB1 variants observed in 60,706 exomes identifies 197 variants that have enough potential to disrupt splicing to warrant further consideration. PMID: 28780672
- AR also indirectly increases the expression of DNA replication genes through stimulatory effects on other metabolic genes with subsequent CDK activation and Rb hyperphosphorylation. PMID: 27760327
- Rb gene promoter methylation was more frequent in gastric cancer patients than in controls. PMID: 28319413
- We report the significance of genetic testing in the early detection and management of retinoblastoma from India. PMID: 26914665
- Results show that the functional state of protein Rb is inferred to be inactive due to its phosphorylation status in the MYCN-amplified retinoblastoma without coding sequence mutations. This makes inactivation of RB1 by gene mutation or by protein phosphorylation a necessary condition for initiating retinoblastoma tumorigenesis, independent of MYCN amplification. PMID: 28211617
- Low RB expression is associated with osteosarcoma. PMID: 28655788
- Loss of RB1 is associated with papillomavirus involvement in Barrett's dysplasia and esophageal adenocarcinoma. PMID: 28722212
- The epigenetic interaction between Linc00441 and bidirectional transcripted neighbor RB1 may be a de novo theory cutting-point for the inactivation of RB1 in hepatocellular carcinoma (HCC). PMID: 28300839
- The data indicate that MAZ is essential to bypass MYB promoter repression by RB family members and to induce MYB expression. PMID: 28973440
- RB inactivation enhances pro-inflammatory signaling through stimulation of the interleukin-6/STAT3 pathway, which directly promotes various malignant features of cancer cells. [review] PMID: 28865172
Q&A
How can I confirm the specificity of a Phospho-RB1 (Ser807/811) antibody in my experimental system?
To confirm specificity of your Phospho-RB1 (Ser807/811) antibody, implement these methodological controls:
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Phosphatase treatment: Treat half of your samples with lambda phosphatase and compare antibody reactivity between treated and untreated samples. Complete signal loss in phosphatase-treated samples confirms phospho-specificity .
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Peptide competition assay: Pre-incubate the antibody with the immunizing phosphopeptide (corresponding to Ser807/811) and observe complete blocking of signal, while pre-incubation with non-phosphorylated peptide should not affect detection .
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Phosphorylation modulation: Compare antibody reactivity in samples with expected phosphorylation differences, such as serum-starved versus serum-stimulated cells, or treatment with CDK inhibitors like Palbociclib, which should show dose-dependent decrease in Ser807/811 phosphorylation .
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Western blot molecular weight verification: Confirm that the detected band appears at the expected molecular weight (approximately 110 kDa) .
Which applications are most commonly supported by Phospho-RB1 (Ser807/811) antibodies?
Phospho-RB1 (Ser807/811) antibodies support multiple experimental applications with specific optimization requirements:
The choice of application should align with your specific research questions, with Western blotting being the most widely validated method for detecting phosphorylation status changes in response to treatments or cellular conditions.
How should I prepare cell lysates to maximize detection of Phospho-RB1 (Ser807/811)?
Optimal cell lysate preparation for Phospho-RB1 (Ser807/811) detection requires careful protocol execution to preserve phosphorylation status:
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Phosphatase inhibition: Include comprehensive phosphatase inhibitor cocktail in lysis buffer to prevent dephosphorylation during sample processing.
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Cell density considerations: For adherent cell lines, aim for 70-80% confluency at harvest to maintain consistent cell cycle distribution and phosphorylation patterns .
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Lysis buffer composition: Use a buffer containing 1% NP-40 or Triton X-100, 150 mM NaCl, 50 mM Tris-HCl (pH 7.4), 1 mM EDTA, 1 mM EGTA, supplemented with protease inhibitors.
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Sample processing: Maintain samples at 4°C throughout processing and avoid repeated freeze-thaw cycles.
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Cell number standardization: For quantitative assays like AlphaLISA, standardize to approximately 2,000-20,000 cells per data point depending on the cell type and expression level .
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Treatment conditions: For positive controls, treat cells with agents that enhance RB1 phosphorylation (e.g., nocodazole, 20% FBS) or decrease phosphorylation (e.g., Palbociclib) to validate antibody responsiveness .
What are the best practices for studying Phospho-RB1 (Ser807/811) in the context of cell cycle regulation?
When investigating Phospho-RB1 (Ser807/811) in cell cycle studies, implement these methodological approaches:
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Cell synchronization: Synchronize cells using serum starvation (G0/G1), double thymidine block (G1/S boundary), or nocodazole treatment (G2/M) to examine phosphorylation changes at specific cell cycle phases .
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CDK inhibitor studies: Use specific CDK4/6 inhibitors like Palbociclib to demonstrate dose-dependent decrease in Ser807/811 phosphorylation, correlating with G1 arrest .
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Multiparameter flow cytometry: Combine Phospho-RB1 (Ser807/811) antibody staining with DNA content analysis using propidium iodide to correlate phosphorylation status with specific cell cycle phases .
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Time-course experiments: Following synchronization release, collect samples at defined time points to track temporal changes in phosphorylation status.
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Correlation with other cell cycle markers: Simultaneously assess cyclin D1, E2F1 target gene expression, and additional RB1 phosphorylation sites to build a comprehensive profile of cell cycle regulation.
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Growth factor stimulation: Compare phosphorylation in serum-starved versus growth factor stimulated conditions (e.g., 20% FBS treatment) to demonstrate cell cycle entry-dependent phosphorylation events .
How can I differentiate between Ser807 and Ser811 phosphorylation and their respective functional significance?
Distinguishing between Ser807 and Ser811 phosphorylation requires sophisticated experimental approaches:
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Site-specific antibodies: While many commercial antibodies detect both Ser807/811 phosphorylation, some are specifically designed to recognize only Ser807 phosphorylation . Compare results using both types of antibodies to identify site-specific patterns.
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Mass spectrometry analysis: For definitive site identification and quantification, perform phosphopeptide enrichment followed by LC-MS/MS analysis to precisely map phosphorylation sites and their relative stoichiometry.
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Site-directed mutagenesis: Generate Ser807→Ala and Ser811→Ala single and double mutants in expression constructs to dissect the functional contribution of each site in cellular models.
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Kinase specificity assays: Different kinases may preferentially target Ser807 versus Ser811. In vitro kinase assays with purified CDKs and site-specific phospho-antibodies can help delineate these specificities.
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Protein-protein interaction studies: Research indicates that Ser807 phosphorylation specifically regulates RB1's association with Bax and its role in apoptosis . Immunoprecipitation experiments comparing wild-type and phospho-site mutants can reveal site-specific interaction partners.
What strategies can address the challenges in detecting Phospho-RB1 (Ser807/811) in tissue samples?
Detecting Phospho-RB1 (Ser807/811) in tissue samples presents unique challenges requiring specialized approaches:
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Tissue fixation optimization: Phospho-epitopes are sensitive to fixation conditions. For paraffin-embedded tissues, limit fixation time in 10% neutral buffered formalin to 24 hours and perform prompt processing to preserve phosphorylation .
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Antigen retrieval methods: Compare heat-induced epitope retrieval using citrate buffer (pH 6.0) versus EDTA buffer (pH 9.0) to determine optimal phospho-epitope exposure conditions .
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Signal amplification systems: For tissues with low expression, employ tyramide signal amplification or polymer-based detection systems to enhance sensitivity while maintaining specificity.
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Phosphatase inhibition during processing: When possible, include phosphatase inhibitors in the tissue collection and processing buffers, particularly for fresh frozen samples.
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Multi-antibody validation: Confirm phosphorylation status using antibodies from different vendors or those recognizing distinct epitopes on phosphorylated RB1 .
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Comparison with proliferation markers: Co-stain with proliferation markers (Ki-67, PCNA) to correlate Phospho-RB1 (Ser807/811) positivity with proliferative status in tissue contexts .
What approaches can reveal the relationship between Phospho-RB1 (Ser807/811) and apoptotic pathways?
Recent research has identified a novel connection between Phospho-RB1 (Ser807/811) and apoptotic regulation through Bax interaction . To investigate this relationship:
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Co-immunoprecipitation studies: Use Phospho-RB1 (Ser807/811) specific antibodies for immunoprecipitation followed by Bax detection, or vice versa, to confirm their interaction under various cellular conditions .
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GST pull-down assays: Employ GST-Bax fusion proteins to pull down interacting proteins from cell lysates, followed by immunoblotting with Phospho-RB1 (Ser807/811) antibodies to validate the specific phosphorylation status of interacting RB1 .
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Apoptosis induction experiments: Compare Phospho-RB1 (Ser807/811) levels and Bax interaction before and after apoptotic stimuli to determine whether this phosphorylation is altered during programmed cell death.
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Subcellular fractionation: Examine the localization of Phospho-RB1 (Ser807/811) in nuclear versus mitochondrial fractions, particularly following apoptotic stimuli, to understand its potential translocation and function at mitochondria where Bax operates.
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Phospho-mimetic mutants: Generate and express S807D/S811D phospho-mimetic RB1 mutants to assess their impact on Bax interaction and apoptotic susceptibility independently of upstream kinase regulation.
How should I properly quantify and normalize Phospho-RB1 (Ser807/811) levels in experimental samples?
Accurate quantification of Phospho-RB1 (Ser807/811) requires careful normalization approaches:
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Total RB1 normalization: Always measure total RB1 protein levels in parallel with phospho-specific detection to calculate the phospho-to-total ratio, accounting for variations in total protein expression .
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Loading control standards: In addition to total RB1, include stable housekeeping proteins (β-actin, GAPDH) or total protein staining (Ponceau S, REVERT) for secondary normalization.
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Standard curve generation: For quantitative assays like AlphaLISA, prepare standard curves using recombinant phosphorylated RB1 protein or phosphopeptides to enable absolute quantification .
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Cell number standardization: Particularly important for flow cytometry and ELISA/AlphaLISA approaches, where signal should be normalized to cell input (typically 2,000-20,000 cells per data point) .
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Signal quantification methods: For Western blots, use digital image analysis with defined background subtraction and avoid saturated signals that prevent accurate quantification.
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Technical and biological replicates: Include at least three technical replicates for each biological sample and multiple biological replicates to ensure reproducibility and enable statistical analysis.
What experimental considerations are important when comparing multiple phosphorylation sites on RB1?
RB1 contains multiple phosphorylation sites that function in concert to regulate its activity. When comparing Ser807/811 with other phosphorylation sites:
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Temporal dynamics analysis: Different sites may be phosphorylated with distinct kinetics following stimulation. Perform detailed time-course experiments to capture these differences .
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Kinase inhibitor profiling: Different CDKs preferentially phosphorylate specific sites. Use selective CDK inhibitors (Palbociclib for CDK4/6, Dinaciclib for CDK1/2) to identify kinase-site relationships .
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Antibody validation controls: Each phospho-specific antibody requires independent validation. Include phosphatase-treated controls for each antibody to confirm specificity .
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Single versus multiple site detection: Consider using antibodies that detect single phosphorylation sites (e.g., Ser807 only) versus those detecting multiple sites (Ser807/811) to dissect site-specific functions .
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Combinatorial phosphorylation analysis: Assess whether phosphorylation at one site influences modification at other sites using sequential immunoprecipitation experiments.
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Functional correlation: Different phosphorylation sites may correlate with distinct RB1 functions. For example, Ser807/811 phosphorylation appears particularly important for Bax interaction and apoptotic regulation, while other sites may more strongly influence E2F repression .
How can Phospho-RB1 (Ser807/811) antibodies be utilized in cancer research and potential therapeutic development?
Phospho-RB1 (Ser807/811) antibodies serve as valuable tools in cancer research applications:
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Biomarker development: Phospho-RB1 (Ser807/811) levels can serve as biomarkers for CDK4/6 inhibitor response in clinical samples. Standardized immunohistochemistry protocols using validated antibodies enable consistent assessment across patient cohorts .
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Drug efficacy monitoring: Track changes in Phospho-RB1 (Ser807/811) levels to assess CDK4/6 inhibitor efficacy in both preclinical models and patient samples. The AlphaLISA assay format provides quantitative measurement capability for therapeutic monitoring .
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Resistance mechanism investigation: In CDK4/6 inhibitor-resistant models, assess Phospho-RB1 (Ser807/811) status to determine whether resistance occurs through RB1-dependent or independent mechanisms.
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Combination therapy rationale: Identify pathways that maintain Phospho-RB1 (Ser807/811) despite CDK4/6 inhibition to develop rationally designed combination therapies targeting these compensatory mechanisms.
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Cancer subtype characterization: Analyze Phospho-RB1 (Ser807/811) patterns across cancer subtypes to identify differential RB1 regulation that might inform subtype-specific therapeutic approaches .
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Patient stratification strategies: Develop quantitative cutoffs for Phospho-RB1 (Ser807/811) positivity that correlate with treatment response to enable prospective patient selection for CDK4/6 inhibitor therapy.
What are the considerations for studying Phospho-RB1 (Ser807/811) in the context of cell differentiation and development?
Beyond cell cycle regulation, RB1 plays important roles in differentiation and development that may involve Ser807/811 phosphorylation:
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Differentiation model systems: Track Phospho-RB1 (Ser807/811) during in vitro differentiation protocols (e.g., myogenesis, adipogenesis, neuronal differentiation) to correlate phosphorylation state with differentiation stage.
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Developmental timing analysis: In developmental studies, precise sampling timing is critical as phosphorylation states may change rapidly during key developmental transitions.
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Tissue-specific expression patterns: Different tissues may exhibit distinctive patterns of Phospho-RB1 (Ser807/811) during development. Immunohistochemistry with phospho-specific antibodies can map these patterns in developmental tissue sections .
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Stem cell studies: Compare Phospho-RB1 (Ser807/811) levels between stem cells and their differentiated progeny to understand the role of this modification in maintaining stemness versus promoting differentiation.
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Transcription factor interactions: During differentiation, RB1 interacts with tissue-specific transcription factors beyond E2F. Co-immunoprecipitation with Phospho-RB1 (Ser807/811) antibodies can identify phosphorylation-dependent interaction partners in differentiation contexts .
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Developmental signaling pathway integration: Investigate how developmental signaling pathways (Wnt, Notch, BMP) influence Phospho-RB1 (Ser807/811) levels to integrate cell cycle control with differentiation programs.
What technical advances are enhancing the detection and analysis of Phospho-RB1 (Ser807/811)?
Recent technological developments are expanding the capabilities for Phospho-RB1 (Ser807/811) analysis:
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Recombinant monoclonal antibody technology: Next-generation recombinant monoclonal antibodies like ZooMAb® provide superior lot-to-lot consistency and specificity compared to traditional antibodies .
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Multiplexed detection platforms: AlphaLISA and similar technologies enable simultaneous quantification of multiple RB1 phosphorylation sites and related proteins from limited sample amounts .
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Single-cell phosphoprotein analysis: Advanced flow cytometry and mass cytometry (CyTOF) techniques allow analysis of Phospho-RB1 (Ser807/811) at the single-cell level, revealing heterogeneity within populations.
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Phosphoproteomic integration: Large-scale phosphoproteomic studies can place Phospho-RB1 (Ser807/811) in the context of the broader phosphorylation landscape, revealing co-regulated phosphorylation events .
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Advanced imaging techniques: Super-resolution microscopy combined with phospho-specific antibodies enables precise subcellular localization of Phospho-RB1 (Ser807/811) and co-localization with interaction partners.
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CRISPR-mediated phosphosite mutation: CRISPR/Cas9 genome editing to introduce specific mutations at Ser807/811 in endogenous RB1 provides more physiologically relevant models than overexpression systems.
