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Target Full Name: Abelson murine leukemia viral oncogene homologue 1

Target Alias: Abelson murine leukemia viral oncogene 1; Abl; C-Abl; JTK7; P150; v-abl Abelson murine leukemia viral oncogene 1; ENSG00000097007; Q13688; Q13914; Q59FK4

Product Type Specific: Abl protein kinase phosphosite-specific antibody

Antibody Code: PK506

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: Y257

Protein UniProt: P00519

Protein SigNET: Abl1

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human Abl (Abl1) sequence peptide Cat. No.: PE-04AFL95

Antibody Immunogen Sequence: GEV(pY)EGV(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues G254 to V260. Y257 phosphorylation is predicted to be inhibitory for phosphotransferase activity. This phosphosite is located in the kinase subdomain I region, which is involved in binding the phosphates in the ATP substrate. This is one of the major in vivo phosphorylation sites in Abl1 (≥335 reports based on mass spectrometry reports recorded in PhosphoSitePlus). It is proposed to regulate intracellular location. Abl is known to be phosphorylated at this site in vitro by Abl1.

Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.

Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.

Antibody Concentration: 1 mg/ml

Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol

Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.

Product Use: Western blotting | Antibody microarray

Antibody Dilution Recommended: 2 µg/ml for immunoblotting

Antibody Potency: Very weak immunoreactivity of a target-sized protein by Western blotting in FBS-treated A431 cells. Medium immunoreactivity with immunogen peptide on dot blots. Very strong immunoreactivity with recombinant human Abl1 on protein dot blots.

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 140-142 kDa.

Antibody Specificity: Very high

Antibody Cross Reactivity: Strong immunoreactivity on protein dot blots with recombinant human Abl2, which features a nearly identical phosphosite sequence. No cross-reactive proteins detected in HEK-293 and A431 cells.

Scientific Background: Abl1 is a protein-tyrosine kinase of the TK group and Abl family. This kinase is highly expressed and widely distributed in most tested human tissues and found in the cytoplasm and nucleus of cells. It has been implicated in the control of a wide range of cellular processes, including cell differentiation, cell division, cell adhesion and stress responses. Abl1 is activated by autophosphorylation as well as by Src-family kinase-mediated phosphorylation. The normal active form of Abl1 is nuclear, and it is sequestered into the cytoplasm by interaction with 14-3-3 through Thr-735 phosphorylation. It localizes to the mitochondria under conditions of oxidative stress. It is a known oncoprotein (OP), although the wild-type form of Abl1 may function as a tumour-suppressor protein (TSP). The DNA-binding activity of Abl1 is regulated by CDK1-mediated phosphorylation. Some cancer-related mutations in human tumours point to a gain of function of the protein kinase. However, the most common mutations (Thr-315 is near kinase Subdomain III; Gly-250 and Glu-255 are located around the ATP-binding kinase Subdomain I) are clustered within the kinase catalytic domain. It is likely that these common mutations may actually inactive Abl1's catalytic activity and reduce its tumour suppressing activity. The active form of the kinase normally acts to inhibit cell proliferation. Translocation t(9;22)(q34;q11) result in chimeric proteins from BCR and Abl1, and this is a cause of chronic myeloid leukemia (CML). BCR-Abl fusion protein results in constitutively active phosphotransferase activity by inhibiting 3BP1/Abl interaction. BCR-Abl1 is found in the cytoplasm and nucleus. BCR-Abl1 induces the Ras, PI3K, and Myc driven cell proliferation pathways. The abnormal phosphorylation of cytoplasmic proteins and receptors in the plasma membrane may result in a gain of function of Abl1 to stimulate cell proliferation. Abl1 is also activated by RIN1 binding to the SH2 and SH3 domains in Abl1, which normally stabilize it in its inactive form. Deletion of the SH3 domain of Abl turns it into an oncogene. Insertional mutagenesis studies in mice also support a role for this protein kinase in mouse cancer oncogenesis. Inhibition of Src/Abl rescued the degeneration of iPSC-derived motor neurons from patients with familial ALS mutations in SOD1, C9orf72, TARDBP, and from sporadic ALS patients (1). This description may include information annotated by UniProt and/or Google AI.

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