Kinexus Products

Target Full Name: Janus protein-tyrosine kinase 2

Target Alias: JAK-2; Janus kinase 2; JTK10; Kinase JAK2 ; JAK2_ENST00000381652; JAK2_ENST00000381652; CCDS6457; A8K910; A9QVL8; B4DYV1; Q506Q0; Q52R86; Q8IXP2; ENSG00000096968

Product Type Specific: JAK2 protein kinase phosphosite-specific antibody

Antibody Code: PK667

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: Y1007+Y1008

Protein UniProt: O60674

Protein SigNET: JAK2

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human JAK2 sequence peptide Cat. No.: PE-04ABZ95

Antibody Immunogen Sequence: KE(pY)(pY)KVKE(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues K1005 to E1012; in the protein kinase catalytic domain activation T loop region between subdomains VII and VIII. T1007 and Y1008 phosphorylation stimulate phosphotransferase activity and induce interactions with SOCS1. These are two of the major in vivo phosphorylation sites in JAK2 (≥19 and ≥18 reports, respectively, from high throughput mass spectrometry studies recorded in PhosphoSitePlus). JAK2 is known to be phosphorylated in vitro at Y1007 by JAK2, and PYK2 (PTK2B, FAK2), and at Y1008 by Abl1, JAK2, and PYK2 (PTK2B, FAK2).

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: 0.5 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 MCF7 cells. Medium immunoreactivity with immunogen peptide on dot blots. Very strong immunoreactivity with recombinant human JAK2 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 125-135 kDa.

Antibody Specificity: Medium

Antibody Cross Reactivity: Strongly immunoreactivity on protein dot blots with recombinant human JAK3 and TYK2, which feature similar phosphosite sequences. No significant cross-reactive proteins detected in phenylarsine oxide (PAO)+vanadate-treated HeLa cells, EGF-treated A431 cells and insulin-treated MCF7 cells, when these cells were homogenized in SDS-PAGE sample buffer. A strong background >75 kDa in insulin-treated MCF7 cells was observed.

Scientific Background: JAK2 is a protein-tyrosine kinase of the TK group and JAKA family. It is a non-receptor kinase involved in the regulation of various cellular processes, including cell cycle progression, development, differentiation, and histone modifications. Additionally, JAK2 plays a critical role in the signalling events of both innate and adaptive immunity as well as erythropoiesis. It features a FERM domain (receptor interaction), an SH2 domain (STAT binding), a pseudokinase domain (negative regulation), and a C-terminal tyrosine kinase domain (catalytic activity). The pseudokinase domain (JH2) normally acts as a brake, preventing the kinase domain from being constantly active. JAK2 can pair with type I receptors to mediate signalling for GHR (growth hormone), PRLR (prolactin), (LEPR) leptin, EPOR (erythropoietin), and THPO (thrombopoietin). JAK2 can also pair with type II receptors to mediate the effects of IFN-alpha, IFN-beta, IFN-gamma, and various interleukins. It is activated by phosphorylation at Tyr-613, and Tyr-972, and Tyr-1008. JAK2 signalling has been implicated in in a wide range of human hematologic malignancies, and this is typically accompanied by the over-activation of the STAT5 transcription factor, which is a substrate. Polycythemia vera, thrombocythemia, and myelofibrosis are classified as clonal myeloproliferative diseases that arise from a multipotent progenitor cell type. These often rare diseases are characterized by excess production of abnormal cells, displacing healthy cells. JAK2 is a known oncoprotein (OP). Cancer-related mutations in human tumours point to a gain of function of the protein kinase. A high percentage of patients with these myeloproliferative diseases have a dominant gain-of-function substitution mutation, V617F, in the JAK2 gene sequence. For example, the V617F mutation was observed in 40 of 45 polycythemia vera patients. The mutation was observed to cause constitutive tyrosine phosphotransferase activity that promoted erythrocytosis in a mouse model. The JAK2 V617F mutation is in the JH2 domain and causes the protein to be permanently turned on, preventing the cell from "turning off" signaling, leading to overproduction of blood cells. This description may include information annotated by UniProt and/or Google AI.

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