PPP2R2A Antibody, Biotin conjugated
Description
Overview of PPP2R2A Antibody, Biotin Conjugated
The PPP2R2A antibody, conjugated with biotin, is a rabbit-derived polyclonal antibody designed to detect the PPP2R2A protein—a regulatory subunit of the serine/threonine protein phosphatase 2A (PP2A) complex. This antibody is widely used in molecular biology research to study PP2A’s role in cellular signaling, tumor suppression, and barrier function. The biotin conjugation facilitates detection in assays such as ELISA, immunohistochemistry (IHC), and immunofluorescence (IF), leveraging streptavidin-based detection systems .
Role in Cellular Signaling
PPP2R2A regulates PP2A activity, which dephosphorylates key signaling proteins like Akt and Jun. Studies using this antibody have shown:
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Tumor Suppression: PPP2R2A inhibits cancer cell proliferation (e.g., prostate cancer) by dephosphorylating oncogenic kinases .
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Barrier Function: It interacts with Jun and filaggrin to maintain epidermal barrier integrity, with knockdown leading to hyperphosphorylation and barrier dysfunction .
Disease Implications
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Cancer: Downregulation of PPP2R2A correlates with metastasis in pancreatic and non-small cell lung cancers .
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Neurodegeneration: PPP2R2A modulates tau phosphorylation, a hallmark of Alzheimer’s disease, suggesting its role in neuroprotection .
Assay Validation
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ELISA: Biotin-conjugated antibodies enable quantification of PPP2R2A levels in cell lysates or tissue homogenates .
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IHC: Paraffin-embedded tissues (e.g., mouse brain) show nuclear and cytoplasmic staining patterns .
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IF: Colocalization with PP2A catalytic subunits confirms complex formation in subcellular compartments .
Technical Considerations
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Research suggests that SAM Domain and HD Domain-Containing Protein 1 (SAMHD1) is specifically targeted by protein phosphatase 2 regulatory subunit Balpha protein (PP2A-B55alpha) holoenzymes during mitotic exit. PMID: 29884836
- Studies indicate that miR614 promotes cell proliferation and inhibits apoptosis of ovarian cancer cells by targeting PPP2R2A. This suggests a potential therapeutic target for ovarian cancer. PMID: 29532877
- These findings provide a novel understanding of the role of PP2A-B55alpha as a crucial factor in meiotic and embryonic development at the onset of life. PMID: 28439046
- Research proposes that the Smad4-Pitx2-PPP2R2A axis, a newly identified signaling pathway, suppresses pancreatic carcinogenesis. PMID: 26848620
- Evidence reveals B55alpha as a PHD2 substrate and highlights a role for PHD2-B55alpha in the response to nutrient deprivation. PMID: 28329677
- Research suggests that PR55alpha promotes pancreatic cancer development by sustaining hyperactivity of multiple oncogenic signaling pathways, including AKT, ERK, and Wnt. PMID: 26893480
- miR-222 targets protein phosphatase 2A subunit B in bladder cancer cells, indicating a potential role in bladder cancer progression. PMID: 26800397
- miR-556-5p functions as an onco-miRNA and contributes to prostate cancer carcinogenesis by suppressing PPP2R2A expression. PMID: 26297546
- Data indicates that breast cancer with PPP2R2A deletions are associated with worse overall survival. Furthermore, the combination of altered PPP2R2A and high CCND1 expression identifies a subgroup of luminal-like breast cancer patients with a high risk of relapse and death. PMID: 25879784
- Overexpression of miR-892a may promote selective growth of colorectal cancer cells by direct suppression of PPP2R2A expression. PMID: 26054685
- miR-136 may play a significant role during TGF-beta1-induced proliferation arrest by targeting PPP2R2A in keratinocytes. PMID: 25654102
- Research suggests that livers of biliary atresia subjects exhibit overexpression of MIR222 (microRNA 222), potentially contributing to liver fibrosis and cell proliferation by targeting PPP2R2A (protein phosphatase 2A subunit B) and Akt signaling. PMID: 25238119
- Suppression of B55alpha activates signaling pathways that may support leukemia cell survival. PMID: 24858343
- Identification of the adenovirus E4orf4 protein binding site on the B55alpha and Cdc55 regulatory subunits of PP2A provides insights into PP2A regulation. PMID: 24244166
- miR-136 promotes Erk1/2 phosphorylation by targeting PPP2R2A in NSCLC cells, suggesting a potential therapeutic target for non-small cell lung cancer treatment. PMID: 23959478
- PPP2R2A status may serve as a predictive marker for the therapeutic efficacy of PARP inhibition. PMID: 23087057
- Both B55alpha and nuclear forkhead box O1 protein (FOXO1) levels are elevated under hyperglycemic conditions in transgenic db/db mouse islets, an animal model of type 2 diabetes. PMID: 22417654
- Data demonstrates that B55alpha antagonizes Cyclin A/Cdk-dependent activation of FoxM1, ensuring that FoxM1 activity is restricted to the G(2) phase of the cell cycle. PMID: 21813648
- Somatic deletions, rather than germline sequence variants of PPP2R2A, may play a more significant role in prostate cancer susceptibility. PMID: 21872824
- The interaction of human adenovirus E4orf4 with both Cdc55 and B55 involves residues within blades 1 and 2, suggesting that E4orf4 binding may block the dephosphorylation of certain PP2A substrates. PMID: 21047956
- Cyclin G2 also associates with various PP2A B' regulatory subunits, as previously shown for cyclin G1. PMID: 11956189
- CFTR is regulated by a direct interaction with the protein phosphatase 2A. PMID: 16239222
- PP2A ABalphaC and ABdeltaC holoenzymes function as positive regulators of Raf1-MEK1/2-ERK1/2 signaling by targeting Raf1. PMID: 16239230
- hAR is a direct target of LEF-1/TCF transcriptional regulation in PCa cells. Expression of the hAR protein is suppressed by a degradation pathway regulated by cross-talk of Wnt, Akt, and PP2A. PMID: 16474850
- Bub1 targets PP2A to centromeres, which in turn maintains Sgo1 at centromeres by counteracting Plk1-mediated chromosome removal of Sgo1. PMID: 16580887
- Deregulation of CHEK2 and/or PPP2R2A is of pathogenic importance in at least a subset of germ cell tumors. PMID: 16790090
- The scaffolding subunit of PP2A exhibits considerable conformational flexibility, which is proposed to play a crucial role in PP2A function. PMID: 17055435
- PP2A binding to Sprouty2 and phosphorylation changes are prerequisites for ERK inhibition downstream of FGFR stimulation. PMID: 17255109
- Protein phosphatase 2A and separase form a complex regulated by separase autocleavage. PMID: 17604273
- The protein phosphatase 2A regulatory subunit alpha4 has a novel role in regulating cell spreading and migration. PMID: 17693407
- PP2A constitutively dephosphorylates the class IIa member HDAC7 to control its biological functions as a regulator of T cell apoptosis and endothelial cell functions. PMID: 18339811
- Homozygous deletions in several biologically important genes, including PPP2R2A and BNIP3L, have been identified in prostate cancer cell lines. PMID: 18670647
- These highly related members of the same subfamily of PP2A regulatory subunits differentially regulate TGF-beta/Activin/Nodal signaling to elicit opposing biological outcomes. PMID: 18697906
- Research shows that E4orf4 protein interacts uniquely with B55 family subunits, and cell killing increases with the level of E4orf4 expression. PMID: 19535438
- Findings suggest that PR55 alpha specifically regulates PP2A-mediated beta-catenin dephosphorylation and plays a crucial role in Wnt signaling. PMID: 19556239
Q&A
What is PPP2R2A and what is its functional significance?
PPP2R2A, also known as serine/threonine-protein phosphatase 2A 55 kDa regulatory subunit B alpha isoform, is a critical component of the PP2A holoenzyme complex. It functions as a regulatory B subunit that modulates substrate selectivity and catalytic activity of the phosphatase . PP2A itself is a major serine/threonine phosphatase involved in numerous cellular processes including signal transduction, cell cycle regulation, and metabolism. The B regulatory subunit (of which PPP2R2A is a member) is particularly important as it determines substrate specificity and subcellular localization of the PP2A holoenzyme . PPP2R2A has multiple aliases including PP2A subunit B isoform B55-alpha, PP2A subunit B isoform PR55-alpha, and PP2A subunit B isoform R2-alpha .
What are the primary applications for PPP2R2A antibody, biotin conjugated?
PPP2R2A antibody, biotin conjugated, is primarily used in immunoassay techniques including ELISA and immunohistochemistry. The currently available products are validated for:
This antibody is particularly valuable for researchers in neuroscience and cancer research fields, especially prostate cancer studies . The biotin conjugation enhances signal detection sensitivity when used with streptavidin-based detection systems, making it particularly useful for experiments requiring enhanced signal amplification.
What specimen types can be analyzed with PPP2R2A antibody, biotin conjugated?
The PPP2R2A antibodies described in the search results are specifically reactive to human samples . Researchers should consider this species reactivity limitation when designing experiments. The antibody is raised in rabbits immunized with either recombinant human PPP2R2A protein fragments (amino acids 2-149) or E. coli-derived human PPP2R2A fragments . No cross-reactivity with other species is explicitly mentioned in the search results, suggesting validation studies would be necessary before applying this antibody to non-human experimental systems.
How is PPP2R2A expression altered in cancer, and what are the implications?
Research has demonstrated that PPP2R2A expression is decreased in a subset of primary prostate tumors . Importantly, studies have shown that loss of PP2A function, including reduced expression of PPP2R2A, can contribute to androgen-independent growth in prostate cancer cells . This suggests PPP2R2A may function as a tumor suppressor in certain contexts.
The mechanistic significance of PPP2R2A loss appears distinct from other PP2A components. While loss of the catalytic subunit PPP2CA promotes androgen depletion-resistant growth by sustaining androgen receptor (AR) transcriptional activity, PPP2R2C (another B regulatory subunit) promotes castration-resistant prostate cancer growth without altering AR expression or canonical AR-mediated signaling . This indicates that different PP2A subunits may contribute to cancer progression through distinct molecular mechanisms.
What experimental approaches can be used to study PPP2R2A function in cellular systems?
Based on the methodologies described in the search results, several experimental approaches can be employed to study PPP2R2A:
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Protein expression analysis: Western blotting techniques following polyacrylamide gel electrophoresis can be used to quantify PPP2R2A protein levels in cell lysates .
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Genetic manipulation: Cloning of PPP2R2A into expression vectors (such as pIRES2-AcGFP1) followed by transfection into cell lines enables overexpression studies .
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Immunohistochemistry: PPP2R2A antibody, biotin conjugated can be used for localization studies in tissue sections, particularly in paraffin-embedded specimens .
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Correlation with clinical outcomes: Analysis of PPP2R2A expression levels in patient samples can be correlated with clinical parameters such as disease progression, metastasis, and cancer-specific mortality .
How does loss of PPP2R2A affect cellular signaling pathways?
The search results indicate that altered PP2A function, including changes in PPP2R2A expression, can affect multiple signaling pathways. In prostate cancer, decreased expression of PPP2R2A has been observed, though the specific downstream effects were not detailed in the provided information .
What are the optimal storage conditions for PPP2R2A antibody, biotin conjugated?
For maximum stability and activity retention, PPP2R2A antibody, biotin conjugated should be stored according to the following recommendations:
The antibody is typically supplied in a liquid form with buffer components that may include:
What validation methods should be employed when using PPP2R2A antibody?
When using PPP2R2A antibody, biotin conjugated, researchers should validate the antibody's specificity and performance in their specific experimental conditions. Recommended validation approaches include:
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Positive and negative controls: Include tissues or cell lines known to express or lack PPP2R2A.
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Blocking peptide competition: Pre-incubation of the antibody with the immunizing peptide should abolish specific staining.
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Technical controls: Include isotype controls and secondary antibody-only controls to assess background and non-specific binding.
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Expression verification: Confirm PPP2R2A expression using alternative methods such as RT-PCR or using antibodies targeting different epitopes.
The antibody purification method (>95%, Protein G purified ) suggests high specificity, but validation remains essential for each experimental system.
What is the recommended protocol for using PPP2R2A antibody, biotin conjugated in ELISA?
While specific ELISA protocols are not detailed in the search results, general guidelines for using biotin-conjugated antibodies in ELISA include:
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Sample preparation: Prepare protein lysates from cells or tissues following standard protocols, such as those described for protein lysate preparation in the search results .
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Protein quantification: Determine protein concentration using methods like the Bradford assay to ensure consistent loading .
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Antibody dilution: Prepare working dilutions of the PPP2R2A antibody, biotin conjugated in appropriate buffer (typically PBS with a carrier protein).
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Detection system: Use streptavidin-conjugated enzymes (HRP or AP) for colorimetric detection, or streptavidin-conjugated fluorophores for fluorescent detection.
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Quantification: Measure signal intensity using appropriate instrumentation (plate reader) and compare to standard curves if absolute quantification is required.
How can PPP2R2A antibody be used in cancer research?
PPP2R2A antibody, biotin conjugated, can be valuable in cancer research through several applications:
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Expression profiling: Analyzing PPP2R2A expression levels in different cancer types and stages to identify potential correlations with disease progression.
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Biomarker development: Based on findings that decreased PPP2R2A expression is associated with certain cancers , the antibody can be used in tissue microarray studies to evaluate PPP2R2A as a potential prognostic or predictive biomarker.
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Mechanistic studies: Investigating the role of PPP2R2A in cancer-related signaling pathways, particularly those involved in cell growth, survival, and therapeutic resistance.
The search results specifically highlight prostate cancer research, where PPP2R2A expression changes have been observed in primary tumors . This suggests PPP2R2A antibodies are particularly relevant for studies involving androgen-dependent and castration-resistant prostate cancer models.
What are the implications of PPP2R2A in neuroscience research?
The specific role of PPP2R2A in neuroscience was not extensively detailed in the search results, though PPP2R2A antibody is identified as relevant for neuroscience research applications . Based on general knowledge of PP2A function in neuronal systems, potential research applications could include:
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Neuronal signaling: Investigating PPP2R2A's role in regulating phosphorylation-dependent neuronal signaling pathways.
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Neurodegenerative diseases: Examining potential alterations in PPP2R2A expression or function in models of neurodegenerative disorders.
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Neuronal development: Studying the role of PPP2R2A-containing PP2A holoenzymes in neuronal differentiation, axon guidance, or synapse formation.
Researchers in neuroscience fields should consider PPP2R2A antibody, biotin conjugated, as a tool for these investigations, particularly when enhanced sensitivity is required.
How can researchers optimize immunohistochemistry protocols using PPP2R2A antibody, biotin conjugated?
For optimal results in immunohistochemistry applications, particularly with paraffin-embedded tissues, researchers should consider the following optimization strategies:
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Antigen retrieval: Test different antigen retrieval methods (heat-induced vs. enzymatic) and buffers (citrate vs. EDTA) to maximize epitope accessibility.
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Antibody dilution: Perform titration experiments to determine the optimal concentration that maximizes specific signal while minimizing background.
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Incubation conditions: Optimize primary antibody incubation time and temperature (overnight at 4°C vs. shorter incubations at room temperature).
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Detection system: When using biotin-conjugated antibodies, consider using streptavidin-HRP systems with tyramide signal amplification for enhanced sensitivity.
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Counterstaining: Select appropriate counterstains that allow clear visualization of PPP2R2A immunoreactivity without interfering with the primary staining.
What factors might contribute to inconsistent results when using PPP2R2A antibody?
Several factors can lead to variability in experimental results when using PPP2R2A antibody, biotin conjugated:
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Antibody stability: Biotin-conjugated antibodies may be susceptible to degradation; proper storage and handling are critical.
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Sample preparation: Variations in fixation methods, fixation duration, and tissue processing can affect epitope preservation and accessibility.
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Endogenous biotin: Tissues with high endogenous biotin (liver, kidney, brain) may produce high background when using biotin-conjugated antibodies without proper blocking steps.
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PPP2R2A expression levels: Natural biological variation in PPP2R2A expression between samples, particularly in disease states, may affect signal intensity.
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Post-translational modifications: Changes in PPP2R2A phosphorylation or other modifications may affect antibody recognition depending on the epitope targeted.
