Phospho-ABL1 (Tyr245) Antibody
Product Specs
Target Background
- Findings demonstrate an important role of c-Abl kinase in Runx1-mediated megakaryocyte maturation and platelet formation, providing a potential mechanism for Abl kinase-regulated hematopoiesis. PMID: 29730354
- Data indicate that c-Abl kinase interacts with and phosphorylates YY1 protein, regulating its transcriptional activity. PMID: 29807053
- The significant role of c-Abl kinase in barrier-altering agonists-mediated cytoskeletal biomechanics has been demonstrated. PMID: 29343719
- While we did not find the AIF1L-ETV6 and ABL1-AIF1L fusions in other ETV6-ABL1-positive ALL, functional studies would be needed to establish the biological role of AIF1L-ETV6 and ABL1-AIF1L and to determine whether they contribute to leukemogenesis and/or to the final leukemia phenotype. PMID: 29726059
- Once activated, c-Abl kinase regulated the activity of Vav1, which further affected the Rac1/PAK1/LIMK1/cofilin signaling pathway. PMID: 29058761
- 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 important when making clinical decisions regarding changing therapy for CML patients treated initially with IM. PMID: 28540759
- Patients with the E255K/V mutation have a poor prognosis, regardless of the stage of the disease at detection. PMID: 29464484
- Therefore, EphA4 is an emerging AbetaOs receptor, and the activation of the EphA4/c-Abl axis would explain the synaptic spine alterations found in Alzheimer's disease. PMID: 29378302
- 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 the BCR/ABL fusion gene with a low detection limit down to 23 fM. It exhibited wide linear ranges over seven orders of magnitude and excellent discrimination ability toward the target. PMID: 27577607
- This study combines a chemical rescue approach with quantitative phosphoproteomics to identify targets of Abl and their phosphorylation sites with enhanced temporal resolution. Both known and novel putative substrates are identified, presenting opportunities for studying unanticipated functions of Abl under physiological and pathological conditions. PMID: 29341593
- 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 PRDX2 gene was found. PMID: 29550484
- We identified a novel mutant p53:c-Abl cytoplasmic signaling complex that promotes MDA-MB-231 cell growth and highlights the contextual cues that confer oncogenic activity to c-Abl in breast cancer. PMID: 28661474
- c-Abl/Arg are oncogenic kinases that regulate differential gene expression. PMID: 28555614
- 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
- We identified a novel c-Abl:p53:p21 signaling axis that functions as a powerful suppressor of mammary tumorigenesis and metastatic progression. PMID: 27626309
- 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
- BGB324 does not inhibit BCR-ABL1 and consequently inhibits chronic myeloid leukemia (CML) independent of BCR-ABL1 mutational status. Our data show that Axl inhibition has therapeutic potential in BCR-ABL TKI-sensitive as well as -resistant CML and support the need for clinical trials. PMID: 27856601
- 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
- These findings show that drug-resistance mutations in the Abl RM exert their allosteric effect by promoting the activated state of Abl and not by decreasing the drug affinity for the kinase. PMID: 28945248
- Germline variants in ABL1 cause a syndrome characterized by congenital heart disease, skeletal abnormalities, and failure to thrive. PMID: 28288113
- c-Abl has a critical role in alpha-synuclein-induced neurodegeneration; selective inhibition of c-Abl may be neuroprotective. PMID: 27348587
- We demonstrate that nanopore technology is suitable for employment in the hematology laboratory for detecting BCR-ABL1 kinase domain mutation in Philadelphia-positive leukemias. PMID: 28663031
- 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
- c-Abl promotes TGF-beta-induced SKIP/Smad3 interaction. PMID: 28666867
- Data indicate the feasibility of detecting ABL1 mutations in cerebrospinal fluid (CSF) by next-generation sequencing (NGS) in patients with central nervous system relapse in BCR-ABL1-positive acute lymphoblastic leukemia. PMID: 28451802
- The e13a2 BCR-ABL1 fusion transcript affects the rate, depth, and 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 may help the choice of the best treatment policy. PMID: 28466557
- Normal ABL1 is a tumor suppressor in BCR-ABL1-induced leukemia. Allosteric stimulation of the normal ABL1 kinase activity enhanced the antileukemia effect of ABL1 tyrosine kinase inhibitors. PMID: 26864341
- The ABL family of tyrosine kinases rheostatically enhances IRE1alpha's enzymatic activities, thereby potentiating endoplasmic reticulum stress-induced apoptosis. PMID: 28380378
- 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
- ETV6-ABL1 fusion occurs in both lymphoid and myeloid leukemias; the genomic profile and clinical behavior resemble BCR-ABL1-positive malignancies, including the unfavorable prognosis, particularly of acute leukemias. The poor outcome suggests that treatment with tyrosine kinase inhibitors should be considered for patients with this fusion. PMID: 27229714
- The c-Abl non-receptor kinase phosphorylates DDB1 at residue Tyr-316 to recruit a small regulatory protein, DDA1, leading to increased substrate ubiquitination. PMID: 28087699
- Drug sensitivity profiles of a set of compound mutations in ABL kinase were also presented in this study. Thus, our large-scale computational study provides comprehensive sensitivity/resistance profiles of ABL mutations toward specific kinase inhibitors. PMID: 28475010
- Though our data support the 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
- Inhibition of c-Abl minimizes receptor recycling pathways and results in chaperone-dependent trafficking of the TfR1 to the lysosome for degradation. PMID: 27226592
- 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
- In this review, we examine the evidence to illuminate the molecular mechanism of ABL1 in the progression of gastric cancer (GC) patients with depression and identify new and effective methods for the initial and long-term treatment of GC. PMID: 27666407
- 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
- Results indicate that eIF4B integrates the signals from Pim and PI3K/Akt/mTOR pathways in Abl-expressing leukemic cells. PMID: 26848623
- The notion that regular and close monitoring of MSI2 mRNA levels in chronic myeloid leukemia patients might identify patients at risk of progressing to blast crisis was not supported by this study; an increase in MSI2 transcripts does not precede an increase in BCR-ABL1 mRNA levels. PMID: 27160312
- Collective results lead us to conclude that GADD45alpha modulates curcumin sensitivity through activation of c-Abl > JNK signaling in a mismatch repair-dependent manner. PMID: 26833194
- ABL kinases promote breast cancer osteolytic metastasis by modulating tumor-bone interactions through TAZ and STAT5 signaling. PMID: 26838548
- Mammalian c-Abl plays an important role in steroid hormone receptor-mediated transcription by regulating RBM39. PMID: 27018250
Q&A
What is the biochemical significance of ABL1 phosphorylation at Tyr245?
Tyrosine 245 occupies a critical position in the SH2-kinase linker domain of ABL1 and plays an essential role in regulating kinase activity. This residue is one of two key tyrosine residues required for autophosphorylation-induced activation of ABL1 intrinsic kinase activity. Phosphorylation at Tyr245 is necessary for maximal wild-type ABL1 kinase activity . Structurally, Tyr245 contributes to one of three "linchpins" that maintain ABL1 in an inactive closed state, thereby serving an important auto-inhibitory function . When phosphorylated, this residue disrupts the SH3 domain-based autoinhibitory interactions, enhancing kinase activity and altering intermolecular associations .
How do Phospho-ABL1 (Tyr245) antibodies differ from general ABL1 antibodies?
Phospho-ABL1 (Tyr245) antibodies specifically detect ABL1 protein only when phosphorylated at tyrosine 245, whereas general ABL1 antibodies detect the protein regardless of its phosphorylation status . This specificity allows researchers to monitor the activation state of ABL1 rather than just its expression level. Additionally, different phospho-specific antibodies are designed for distinct experimental applications (WB, IF, ELISA) with varying sensitivities and optimal dilution ranges . When selecting these antibodies, researchers should also consider whether they cross-react with other phosphorylated proteins – for example, some Phospho-ABL1 (Tyr245) antibodies may cross-react with activated EGF receptor .
What are the optimal conditions for detecting phosphorylated ABL1 (Tyr245) in Western blotting experiments?
For optimal Western blot detection of phosphorylated ABL1 at Tyr245, a multifaceted approach is necessary:
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Sample preparation: Cell lysates should ideally come from cells either stimulated with appropriate growth factors or treated with phosphatase inhibitors (e.g., pervanadate) to preserve phosphorylation
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Antibody dilution: Most Phospho-ABL1 (Tyr245) antibodies work optimally at 1:500-1:2000 dilution for Western blotting
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Controls: Include both positive controls (K562 cells that express BCR-ABL1 or pervanadate-treated Jurkat cells) and negative controls (untreated cells)
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Molecular weight identification: Expect to visualize a 145 kDa band corresponding to c-Abl, and potentially a 210 kDa band corresponding to BCR-Abl fusion protein in appropriate cell lines
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Blocking conditions: Use 5% BSA in TBST rather than milk, as milk contains phosphoproteins that may interfere with phospho-specific antibody binding
How can researchers validate the specificity of Phospho-ABL1 (Tyr245) antibody signals?
To validate antibody specificity, employ the following methodological approaches:
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Peptide competition assay: Pre-incubating the antibody with the phosphorylated peptide immunogen should eliminate specific signal, while pre-incubation with non-phosphorylated peptide should not affect binding
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Phosphatase treatment: Treating cell lysates with lambda phosphatase should abolish the signal if it's truly phospho-specific
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Mutant expression: Utilize cells expressing ABL1 with Tyr245 mutated to a non-phosphorylatable residue (e.g., Tyr245Phe) as a negative control
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siRNA knockdown: Reduction of ABL1 expression should correspondingly reduce phospho-ABL1 signal
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Kinase inhibitor treatment: Cells treated with ABL1 kinase inhibitors (e.g., imatinib) should show reduced phosphorylation at Tyr245
How should researchers interpret contradictory findings regarding phosphorylation at Tyr245 in different ABL1 mutants?
The research literature presents an interesting contradiction: while the p.Tyr245Phe substitution reduces ABL1 kinase activity by approximately 50%, the p.Tyr245Cys substitution appears to increase kinase activity . To properly interpret such contradictory findings:
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Consider structural context: Different amino acid substitutions at the same position may have opposing effects based on their chemical properties. The cysteine substitution may disrupt autoinhibition more severely than phenylalanine
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Examine experimental systems: Variations in experimental systems (cell types, expression levels, assay conditions) may contribute to discrepancies
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Analyze potential compensatory mechanisms: Secondary phosphorylation events or conformational changes may compensate differently depending on the specific mutation
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Investigate through multiple approaches: Employ both in vitro kinase assays and cellular phosphorylation studies to confirm findings
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Consider impacts on protein-protein interactions: Different substitutions may differentially affect interactions with regulatory partners like ABI1
What factors can lead to false positives or false negatives when using Phospho-ABL1 (Tyr245) antibodies?
Multiple factors can compromise accurate detection of phosphorylated ABL1:
False positives:
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Cross-reactivity with other phosphorylated proteins, particularly activated EGF receptor
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Inappropriate secondary antibody selection leading to background signals
False negatives:
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Rapid dephosphorylation during sample preparation without adequate phosphatase inhibitors
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Sub-optimal lysis conditions failing to extract nuclear or cytoskeletal-bound ABL1
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Competitive binding from other SH3-domain proteins in complex lysates
How can Phospho-ABL1 (Tyr245) antibodies be used to study developmental disorders associated with ABL1 mutations?
Phospho-ABL1 (Tyr245) antibodies provide valuable tools for investigating developmental disorders linked to ABL1 mutations through several approaches:
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Patient sample analysis: These antibodies can assess phosphorylation levels in patient-derived cells to correlate ABL1 activation with disease phenotype severity
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Disease modeling: In cellular and animal models expressing ABL1 mutations (like p.Tyr245Cys or p.Ala356Thr), these antibodies help monitor altered signaling pathways contributing to congenital heart disease and skeletal abnormalities
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Developmental pathway investigation: They enable researchers to examine the relationship between ABL1 activation and TGF-β signaling pathways during tissue development
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Therapeutic screening: These antibodies can evaluate the efficacy of potential therapeutic compounds designed to normalize phosphorylation levels in cells with pathogenic ABL1 variants
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Genotype-phenotype correlation: By quantifying Tyr245 phosphorylation in different ABL1 mutants, researchers can establish correlations between specific mutations, kinase activity levels, and clinical manifestations
What methodological approaches can determine whether increased phosphorylation at Tyr245 represents a gain or loss of function?
Determining whether increased Tyr245 phosphorylation represents gain or loss of function requires comprehensive methodology:
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Downstream substrate phosphorylation assessment: Measure phosphorylation of known ABL1 substrates (e.g., STAT5) using phospho-specific antibodies against these targets
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Cell-based functional assays: Evaluate cellular processes regulated by ABL1 (cytoskeletal remodeling, endocytosis, DNA damage response) in systems with altered Tyr245 phosphorylation
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In vitro kinase activity assays: Directly measure the catalytic activity of immunoprecipitated ABL1 using synthetic substrates
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Comparative mutation analysis: Compare the effects of different mutations affecting Tyr245 (e.g., p.Tyr245Cys vs. p.Tyr245Phe) on kinase activity and downstream signaling
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Structure-function correlation: Combine phosphorylation data with structural analysis to understand how phosphorylation alters protein conformation and function
What strategies can resolve weak or inconsistent signals when using Phospho-ABL1 (Tyr245) antibodies?
To address weak or inconsistent signals:
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Optimize cell stimulation: Treat cells with pervanadate or appropriate growth factors to maximize Tyr245 phosphorylation
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Refine lysis conditions: Use buffer containing 1% NP-40 or RIPA with phosphatase inhibitors (sodium orthovanadate, sodium fluoride, and β-glycerophosphate) to preserve phosphorylation state
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Adjust antibody concentration: Test a range of antibody dilutions (1:500 to 1:2000) to determine optimal concentration for your specific experimental system
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Enhance detection systems: Consider using more sensitive detection methods such as enhanced chemiluminescence (ECL) substrates or fluorescently-tagged secondary antibodies
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Optimize blocking conditions: Use 5% BSA instead of milk to prevent interference from phosphoproteins in milk
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Consider cellular localization: Remember that ABL1 shuttles between nucleus and cytoplasm, so subcellular fractionation may improve detection of phosphorylated pools
How can researchers distinguish between phosphorylated c-ABL1 and BCR-ABL1 fusion proteins?
Distinguishing between phosphorylated c-ABL1 and BCR-ABL1 requires careful experimental design:
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Molecular weight discrimination: On Western blots, c-ABL1 appears at approximately 145 kDa, while BCR-ABL1 appears at approximately 210 kDa
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Cell line selection: Use appropriate positive controls - K562 cells express BCR-ABL1, while many other cell lines express only c-ABL1
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Isoform-specific antibodies: When necessary, combine Phospho-ABL1 (Tyr245) antibodies with BCR-specific antibodies in co-immunoprecipitation or dual-staining experiments
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Control experiments: Include specific inhibitors like imatinib at concentrations that differentially affect BCR-ABL1 versus c-ABL1
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RNA expression analysis: Correlate protein detection with transcript expression analysis to confirm which form is being detected
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Subcellular localization studies: BCR-ABL1 is predominantly cytoplasmic, while c-ABL1 shuttles between nucleus and cytoplasm
