Product Name:
BARK1-Y356
Product Number:
ab-pk537
Target Full Name: Beta-adrenergic receptor kinase 1
Target Alias: ADRBK1; Adrenergic, beta, receptor kinase 1; ARBK1; BARK; BARK1; Beta-adrenergic receptor kinase 1; Beta-ARK-1; EC 2.7.11.15; FLJ16718; G- protein coupled receptor kinase 2; Kinase GRK2
Product Type Specific: GRK2 protein kinase phosphosite-specific antibody
Antibody Code: PK537
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y356
Protein UniProt: P25098
Protein SigNET: BARK1
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human BARK1 (GRK2, ADRBK1) sequence peptide Cat. No.: PE-04ACJ85
Antibody Immunogen Sequence: THG(pY)MAP(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues T353 to P359; in the protein kinase catalytic domain activation T loop region between subdomains VII and VIII. Y356 phosphorylation is predicted to be stimulatory for phosphotransferase activity. This is one of the major in vivo phosphorylation sites in BARK1 (≥71 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus).
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: Strong immunoreactivity of a target-sized protein by Western blotting in FBS-treated Jurkat cells and weakly in HeLa cells. Very strong immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus Macaque | Dog | Rat | Mouse | Fruit fly | Honey Bee | Nematode worm.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 78-85 kDa.
Antibody Specificity: Very high
Antibody Cross Reactivity: No medium to strong cross-reactivities with other proteins in Jurkat cells, HeLa cells, and sea star oocytes.
Related Product 1: BARK1 (353-359) pY356 antibody blocking peptide (Cat. No.: PE-04ACJ85)
Related Product 2: BARK1 S670 phosphorylation antibody (Cat. No.: AB-PK536)
Scientific Background: BARK1 (GRK2, ADRBK1) is an 80 kDa protein-serine/threonine kinase of the AGC group and GRK family. It features three distinct domains: an N-terminal RGS homology (RH) domain, a central protein kinase domain, and a C-terminal pleckstrin homology (PH) domain, which binds Gb/g subunits.It is activated by G-beta/gamma subunit complex. It specifically phosphorylates the agonist-occupied (active) form of beta-adrenergic receptors and related GPCRs, leading to receptor desensitization and internalization. Besides GPCRs, it phosphorylates other substrates, such as chemokine receptors, cytoskeletal proteins, and those involved in the Hedgehog signaling pathway. Phosphorylation of Ser-29 increases phosphotransferase activity. Phosphorylation of Ser-670 inhibits G-beta/gamma-mediated activation of GRK2. Phosphorylation of Tyr-13, Tyr-86, Tyr-92 and Ser-685 increases interaction with G-alpha(q) and may induce protein degradation. Elevated levels of BARK1 are associated with heart failure, where it disrupts the normal function of beta-adrenergic receptors, contributing to diminished contractility. BARK1 deficiency leads to severe cardiac developmental defects and embryonic lethality in mice. This description may include information annotated by UniProt and/or Google AI.