Phospho-BCR (Y360) Antibody
Product Specs
Target Background
- The combination of BCR-ABL1 transcript type and spleen size at diagnosis is significantly predictive for achieving an overall MMR and FFS. Incorporating these predictors could be vital when making clinical decisions regarding therapy changes for CML patients treated initially with IM. PMID: 28540759
- Expression of WASP inversely correlates with BCR-ABL1 levels and the progression of the disease in Chronic myeloid leukemia patients. BCR-ABL1 downregulates WASP in part by epigenetic modification of its proximal promoter. PMID: 29022901
- The imaging method achieved ultrasensitive detection of BCR/ABL fusion gene with a low detection limit down to 23 fM. This method exhibits wide linear ranges over seven orders of magnitude and excellent discrimination ability toward the target. PMID: 27577607
- This is the first report evaluating the role of SOD2 in native and T351-mutated BCR-ABL-expressing cells and in a large cohort of chronic myeloid leukemia patients. In leukemic cells silenced for SOD2 expression, a specific down-regulation of the expression of the PRDX2 gene was found. PMID: 29550484
- The compound missense mutations in the BCR-ABL kinase domain are responsible for eliciting disease progression, drug resistance, or disease relapse in chronic myeloid leukemia. PMID: 28278078
- JNJ-26854165, an inhibitor of MDM2, inhibits proliferation and triggers cell death in a p53-independent manner in various BCR/ABL-expressing cells, including primary leukemic cells from patients with CML blast crisis and cells expressing the Imatinib-resistant T315I BCR/ABL mutant. PMID: 27999193
- Double inhibition of the N- and C-terminal termini can disrupt Hsp90 chaperone function synergistically, but not antagonistically, in Bcr-Abl-positive human leukemia cells. PMID: 28036294
- This study identifies different BCR/Abl protein suppression patterns as a converging trait of chronic myeloid leukemia cell adaptation to energy restriction. PMID: 27852045
- BCR-ABL1-positive microvesicles from chronic myeloid leukemias malignantly transform human bone marrow mesenchymal stem cells. PMID: 28836580
- Data indicate that the Sp1 oncogene functions as a positive regulator for BCR/ABL expression. PMID: 27144331
- Dehydrocostus lactone significantly inhibits the phosphorylation expression of Bcr/Abl, STAT5, JAK2, and STAT3, and downstream molecules including p-CrkL, Mcl-1, Bcl-XL, and Bcl-2 proteins in K562 cells. PMID: 28300289
- H19 overexpression, a frequent event in chronic myeloid leukemia, was associated with higher BCR-ABL transcript and disease progression. H19 DMR/ICR hypomethylation in CML may be one of the mechanisms mediating H19 overexpression. PMID: 28776669
- Frequent molecular monitoring and intervention are required for patients who do not show a reduction in BCR-ABL1 transcripts to these levels after stem cell transplantation. PMID: 27334764
- This study shows that BCR regulates inflammation development via the alpha subunit of casein kinase II associated with BCR. PMID: 27630163
- The e13a2 BCR-ABL1 fusion transcript affects the rate, the depth, and the speed of the response to treatment with imatinib firstline, and that including the transcript type in the calculation of the baseline risk scores may improve prognostic stratification and help choose the best treatment policy. PMID: 28466557
- Cell division cycle protein 6 overexpression may contribute to the high proliferation and low apoptosis in chronic myeloid leukemia cells and can be regulated by BCR/ABL signal transduction through downstream phosphoinositide 3-kinase/Akt and Janus kinase/signal transducer and activator of transcription pathways, suggesting cell division cycle protein 6 as a potential therapeutic target in chronic myeloid leukemia. PMID: 28639894
- While our data support previous findings that co-expression of BCR-ABL transcripts is due to the occurrence of exonic and intronic polymorphisms in the BCR gene, it also shows that the intronic polymorphism can arise without the linked exonic polymorphism. The occurrence of ABL kinase domain mutation is less frequent in the Indian population. PMID: 27748288
- In silico three-dimensional modeling of apoptin, molecular docking experiments between the apoptin model and the known structure of Bcr-Abl, and the 3D structures of SH2 domains of CrkL and Bcr-Abl, were performed. PMID: 22253690
- The present study screened for the presence of bcr-abl transcripts in the blood of a group of healthy individuals. PMID: 24535287
- Data indicate that the biosensor showed excellent analytical performance for the detection of the BCR/ABL oncogene in clinical samples of patients with leukemia. PMID: 27693719
- Studies indicate that the prognosis of BCR-ABL-positive acute myeloid leukemia (BCR-ABL+ AML) seems to depend on the cytogenetic and/or molecular background rather than on BCR-ABL itself. PMID: 27297971
- Depletion of endogenous MAPK15 expression inhibited BCR-ABL1-dependent cell proliferation. PMID: 26291129
- This study identifies a novel BCR-ABL/IkappaBalpha/p53 network, whereby BCR-ABL functionally inactivates a key tumor suppressor in chronic myeloid leukemia. PMID: 26295305
- Blockade of the interaction between Bcr-Abl and PTB1B by small molecule SBF-1 overcomes imatinib-resistance of K562 cells. PMID: 26721204
- BCR-ABL1 transcript types found in Syria were similar to that of Indian Far-Eastern, African, or European populations. The M-BCR rearrangement types were not dependent on white blood count, platelet count, hemoglobin level, or gender of the patients. PMID: 27273956
- Data suggest that elevated interleukin-1beta secretion from tyrosine kinase inhibitor- (TKI-)resistant chronic myeloid leukemia (CML) cells contributes to TKI/imatinib resistance via promotion of cell viability/migration; cells used lack BCR-ABL mutation. PMID: 26831735
- Higher incidence of BCR-ABL and lower incidence of TEL-AML1 are associated with acute lymphoblastic leukemia. PMID: 26264145
- AIC-47 in combination with imatinib strengthened the attack on cancer energy metabolism in BCR-ABL-harboring leukemic cells. PMID: 26607903
- Molecular rearrangements and the minimal residual disease follow-up for 5 chronic myeloid leukemia patients; 3 resulted from new rearrangements between the BCR and ABL1 sequences (the breakpoints being located within BCR exon 13 in 2 cases and within BCR exon 18 in one case). The other 2 cases revealed a complex e8-[ins]-a2 fusion transcript involving a 3rd partner gene. PMID: 26252834
- Analysis of WT1 and M-BCR-ABL expressions in chronic myeloid leukemia reveals that high WT1 expression in CML patients is detected especially in the advanced stages of the disease. PMID: 26429162
- BCR-ABL kinase domain mutation is associated with Philadelphia-positive leukemia. PMID: 25379619
- Separase protein levels decrease and Separase proteolytic activity increases exclusively in b3a2 p210BCR-ABL-positive cell lines under Imatinib treatment. PMID: 26087013
- Our results confirm that not obtaining a BCR-ABL/ABL ratio of PMID: 25756742
- We describe a regulatory pathway modulating BCR and BCR/ABL1 expression, showing that the BCR promoter is under the transcriptional control of the MYC/MAX heterodimer. PMID: 26179066
- BCR- ABL1 mutations are associated with clinical resistance, but may not be considered the only cause of resistance to imatinib. PMID: 25740611
- Expression profiling of adult acute lymphoblastic leukemia identifies a BCR-ABL1-like subgroup characterized by high non-response and relapse rates. PMID: 25769542
- Expression of the BCR-ABL gene were confirmed by FISH, which revealed a high concordance (100%) rate. We found that real-time RT-qPCR is more reliable and should be used in Moroccan biomedical analysis laboratories to monitor CML progression. PMID: 25730044
- BCR-ABL1 mutation is associated with chronic myeloid leukemia. PMID: 25721898
- Data suggest that the acquisition of additional BCR-ABL1 fusion genes in chronic myeloid leukemia (CML) in the blast phase (BP) through mitotic recombination between the derivative chromosome and the normal homologue. PMID: 26186983
- Lack of response to tyrosine kinase inhibitors associated with mutation in the BCR-ABL gene was significantly higher in imatinib-treated patients, and all mutations arose after treatment. T315I was a common treatment-emergent mutation. PMID: 25615000
- High proportion of M-bcr gene is associated with chronic myeloid leukemia. PMID: 25520136
- ATRA treatment decreased DNA damage repair and suppressed acquisition of BCR-ABL mutations. PMID: 24967705
- Results suggest that the EGFR and Akt pathways are involved in the regulation of BCRP expression in non-small cell lung carcinoma cells. PMID: 25479544
- AIC-47, acting through the PPARgamma/beta-catenin pathway, induced down-regulation of c-Myc, leading to the disruption of the bcr-abl/mTOR/hnRNP signaling pathway, and switching of the expression of PKM2 to PKM1 in acute myeloid leukemia. PMID: 25644089
- Bcr knockdown in the context of KSHV-associated disease might enhance Rac1-mediated angiogenesis. PMID: 25631082
- The epigenetic silencing of miR-23a led to derepression of BCR/ABL expression, and consequently contributes to CML development and progression. PMID: 25213664
- STAT3 is a critical signaling node in BCR-ABL1 tyrosine kinase-independent leukemia resistance that is reversed by a discovered BP-5-087. PMID: 25134459
- C817 is a promising compound for treatment of CML patients with Bcr-Abl kinase domain mutations that confer imatinib resistance. PMID: 24487968
- BCR-ABL-T315I mutation is associated with chronic myeloid leukemia. PMID: 25217883
- Leptin levels were increased in BCR-ABL p210 positive chronic myeloid leukemia patients. PMID: 25648025
Q&A
What is the biological significance of BCR Y360 phosphorylation?
BCR Y360 phosphorylation plays a critical role in regulating BCR's enzymatic activity. Research has demonstrated that Y360 is essential for the transphosphorylation activity of BCR. When phosphorylated at this site (particularly by the Bcr-Abl oncoprotein), BCR's serine/threonine kinase activity is significantly inhibited, impairing both auto- and transkinase activities . This inhibitory mechanism has profound implications in Philadelphia chromosome-positive leukemia, where BCR's normal tumor-suppressive functions may be compromised through this phosphorylation event .
How does phosphorylation at Y360 differ functionally from other BCR phosphorylation sites?
BCR contains multiple phosphorylation sites with distinct functional outcomes:
| Phosphorylation Site | Key Functions | Interacting Partners | Disease Relevance |
|---|---|---|---|
| Y360 | Critical for transphosphorylation activity | Not fully characterized | Inhibited in Ph+ leukemia |
| Y177 | Creates GRB2 binding site; activates RAS pathway | GRB2, SOS1 | Crucial for BCR-ABL leukemogenesis |
| Y283 | Minimal effect on kinase activity | Not well characterized | Less studied |
While Y177 phosphorylation creates a binding site for GRB2 and is highly specific for this interaction , Y360 phosphorylation appears more directly involved in regulating BCR's intrinsic enzymatic functions . Interestingly, mutation studies have shown that Y177F mutants had greatly reduced ability to transphosphorylate casein and histone H1, whereas Y360F mutations had little effect on BCR's autophosphorylation activity but significantly impacted its transphosphorylation capabilities .
What are the validated applications and optimal protocols for using Phospho-BCR (Y360) Antibody?
Current commercial Phospho-BCR (Y360) antibodies have been validated for multiple applications with specific recommended protocols:
| Application | Dilution Range | Key Protocol Considerations |
|---|---|---|
| Western Blotting | 1:500-1:2000 | Use phosphatase inhibitors during sample preparation |
| Immunohistochemistry | 1:100-1:300 | Paraffin-embedded sections show good results |
| Immunofluorescence | 1:200-1:1000 | Fixed/permeabilized specimens yield clearest signals |
| ELISA | 1:40000 | High sensitivity for quantitative analysis |
For Western blotting applications, researchers should anticipate detecting bands at approximately 160 kDa (BCR) and/or 210 kDa (BCR-ABL fusion protein) . When designing experiments, remember that antibodies like Boster Bio's A00022Y360 are generated against synthesized peptides derived from human BCR surrounding the Y360 phosphorylation site (amino acid range 331-380) .
What controls should be incorporated when using Phospho-BCR (Y360) Antibody in experimental designs?
Robust experimental design requires appropriate controls to validate Phospho-BCR (Y360) antibody specificity:
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Phosphopeptide competition assay: Pre-incubating the antibody with synthetic phospho-Y360 peptide should abolish signal in all applications, as demonstrated in validation studies .
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Phosphatase treatment: Treating duplicate samples with lambda phosphatase provides a negative control by removing phosphorylation.
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Kinase inhibitor treatment: For cells expressing BCR-ABL, treatment with tyrosine kinase inhibitors like STI-571 (Gleevec) should reduce Y360 phosphorylation, serving as a biological negative control .
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Y360F mutant expression: Cells expressing BCR with a Y360F mutation provide the most stringent negative control as this residue cannot be phosphorylated.
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MEK1/2 inhibitor treatment: Recent research shows MEK1/2 inhibition significantly reduces Y360 phosphorylation in leukemic cells, offering an additional pathway-specific control .
Implementing these controls systematically ensures confident interpretation of experimental results.
How can Phospho-BCR (Y360) Antibody be utilized to investigate signaling pathway crosstalk?
The Phospho-BCR (Y360) Antibody provides a powerful tool for examining signaling network interactions:
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MEK1/2 pathway interactions: Studies have demonstrated that MEK1/2 inhibition significantly reduces Y360 phosphorylation in leukemic cells, suggesting an important connection between MAPK signaling and BCR regulation .
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Wnt signaling pathway: BCR functions as a negative regulator of Wnt signaling through interactions with β-catenin. Research indicates that tyrosine phosphorylation of BCR (potentially including Y360) may regulate this interaction, particularly in the cytoplasm .
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BCR-ABL core complex analysis: In CML research, ultracentrifugation in sucrose gradients coupled with immunoblotting using Phospho-BCR (Y360) antibody can help characterize the BCR-ABL signaling complex and how it changes with tyrosine kinase inhibitor treatment .
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Phosphorylation network mapping: Combining Phospho-BCR (Y360) detection with antibodies against other phosphorylated proteins can help construct comprehensive phosphorylation networks in normal and disease states .
These approaches facilitate understanding of BCR's position within broader cellular signaling networks.
How does the regulation of BCR Y360 phosphorylation differ between normal and leukemic cells?
The regulation of BCR Y360 phosphorylation exhibits important differences between normal and leukemic contexts:
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Normal cells: In healthy cells, Y360 phosphorylation is likely a regulated event controlled by specific stimuli and signaling pathways, contributing to BCR's normal functions in cellular homeostasis.
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BCR-ABL+ leukemic cells: In Philadelphia chromosome-positive leukemias, the constitutively active BCR-ABL tyrosine kinase causes hyperphosphorylation of BCR at Y360, inhibiting its serine/threonine kinase activity .
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Pathway dependence: In leukemic cells, MEK1/2 inhibition experiments show that Y360 phosphorylation depends partially on MAPK pathway activity, indicating pathway rewiring in the disease state .
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Response to kinase inhibitors: In BCR-ABL+ cells treated with tyrosine kinase inhibitors like STI-571, Y360 phosphorylation decreases, but the complex interplay with other signaling proteins may result in residual phosphorylation through compensatory mechanisms .
Understanding these differences provides insights into leukemogenesis and therapeutic response mechanisms.
What are common technical challenges in detecting BCR Y360 phosphorylation and their solutions?
| Challenge | Possible Causes | Recommended Solutions |
|---|---|---|
| Weak or absent signal | Insufficient phosphorylation; phosphatase activity | Add phosphatase inhibitors to lysis buffer; Enrich phosphoproteins before analysis; Concentrate samples |
| Multiple bands | Cross-reactivity; protein degradation | Validate with phosphopeptide competition; Add protease inhibitors; Optimize antibody dilution |
| Inconsistent results between applications | Context-dependent epitope accessibility | Optimize protocols for each application; Consider native vs. denatured conditions |
| High background | Non-specific binding; Insufficient blocking | Increase blocking time/concentration; Optimize antibody dilution; Use additional wash steps |
For particularly challenging samples, researchers might consider phosphotyrosine enrichment before analysis or using the more sensitive phospho-ELISA approach that has been validated for this antibody .
How can contradictory findings between phospho-specific and total BCR detection be reconciled?
When phospho-specific and total BCR antibodies yield apparently contradictory results, consider these analytical approaches:
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Epitope masking: Y360 phosphorylation may mask epitopes recognized by certain total BCR antibodies, creating an illusion of decreased expression when phosphorylation increases.
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Subcellular redistribution: Phosphorylation at Y360 may alter BCR's localization, resulting in apparent changes in detection depending on fractionation methods. This is supported by findings showing that BCR-β-catenin complex formation is regulated by tyrosine phosphorylation and differs between nuclear and cytoplasmic fractions .
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Turnover rate changes: Phosphorylation status can affect protein stability and degradation rates, leading to actual differences in total protein levels.
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Methodological considerations: Always evaluate whether the lysis conditions preserve phosphorylation states and whether denaturing conditions equally expose epitopes for both antibodies.
For accurate interpretation, normalize phospho-signal to total BCR detected in parallel samples under identical conditions, and validate with orthogonal methods.
How can Phospho-BCR (Y360) Antibody contribute to chronic myeloid leukemia (CML) research?
Phospho-BCR (Y360) Antibody offers unique insights for CML research:
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Therapeutic monitoring: Monitoring changes in Y360 phosphorylation following tyrosine kinase inhibitor treatment can provide early indicators of drug efficacy or developing resistance .
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Disease mechanisms: Investigating how Y360 phosphorylation affects BCR's tumor-suppressive functions helps elucidate fundamental leukemogenesis mechanisms .
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Leukemic stem cell studies: Examining Y360 phosphorylation in primitive leukemic stem cells versus differentiated blasts may reveal signaling differences that contribute to therapy resistance.
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Combination therapy approach: MEK1/2 inhibitors reduce Y360 phosphorylation in leukemic cells , suggesting potential for combination strategies targeting both BCR-ABL and MEK pathways.
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Biomarker development: Y360 phosphorylation patterns could potentially serve as prognostic or predictive biomarkers for CML patient stratification.
These applications demonstrate how mechanistic insights from fundamental phosphorylation studies can translate to clinical research.
What is the emerging role of BCR Y360 phosphorylation in SH2 domain interaction studies?
Recent research has revealed interesting connections between BCR phosphorylation and SH2 domain interactions:
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Binding specificity: While Y177 creates a well-characterized binding site for the GRB2 SH2 domain, the specific SH2 domain-containing proteins that might interact with phosphorylated Y360 remain under investigation .
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SH2 domain modeling: Advanced sequence-to-affinity models for SH2 domains now incorporate multi-round selection data that can be applied to predict potential interactors with phosphorylated Y360 .
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Competitive binding analysis: Fluorescence polarization competition assays using synthetic phosphopeptides containing Y360 can quantify binding affinities of candidate SH2 domain proteins .
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Signaling network implications: The identification of SH2 domain proteins that specifically recognize phospho-Y360 would expand our understanding of how this modification regulates BCR's role in protein-protein interaction networks .
This emerging area highlights how Phospho-BCR (Y360) Antibody can contribute to broader phosphotyrosine signaling research beyond its use in simple detection assays.
