Product Name:
Rock2-pY722
Product Number:
ab-pk799
Target Full Name: Rho-associated protein kinase 2
Target Alias: KIAA0619; Kinase Rock2; P164 Rock-2; RhoA-binding serine/threonine kinase alpha; Rho-associated protein kinase 2; Rho-associated, coiled-coil containing protein kinase 2; Rock-II; ROK-alpha
Product Type Specific: ROCK2 protein kinase phosphosite-specific antibody
Antibody Code: PK799
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y722
Protein UniProt: O75116
Protein SigNET: Rock2
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human Rock2 (ROKa) sequence peptide Cat. No.: PE-04ALB99
Antibody Immunogen Sequence: NKI(pY)ESI(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues N719 to I725; in the region between the kinase catalytic and Rho_Binding domains. Y722 phosphorylation inhibits phosphotransferase activity and inhibits interaction with RhoA. This is the major site of in vivo phosphorylation of ROCK2 based on ≥121 high throughput mass spectrometry reports recorded in PhosphoSitePlus. ROCK2 is known to be phosphorylated at this site in vitro by Src.
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 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 | Rhesus Macaque | Dog | Rat | Mouse | Chicken | Frog.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 160-180 kDa.
Antibody Cross Reactivity: Almost no significant cross-reactivities detected in Jurkat MCF7 cells. In HepG2 cells, a very high background was noted.
Related Product 1: Rock2-pY722 blocking peptide
Related Product 2: Rock2Subtide - ROCK2 protein kinase substrate peptide
Related Product 3: ROCK1 T233 phosphorylation antibody (Cat. No.: AB-PK947)
Related Product 4: Rock1-pY913 phosphosite-specific antibody (Cat. No.: AB-PK798)
Scientific Background: ROCK2 (Rho-associated, coiled-coil containing protein kinase 2; ROK-alpha) is a 160 kDa protein-serine/threonine kinase of the AGC group and DMPK family. It regulates the actin cytoskeleton and cell polarity, thus is involved in the regulation of smooth muscle contraction, formation of stress fibre and focal adhesions, cell adhesion, and cell motility. It is primarily responsible for phosphorylating substrates like myosin light chain (MLC) and LIM kinase, promoting actin stress fiber formation, smooth muscle contraction, and cytokinesis. ROCK2 features a kinase domain at the N-terminus, a coiled-coil region, a Rho-binding domain (RBD), and a pleckstrin homology (PH) domain. It exists in an auto-inhibited form that is activated upon RhoA-GTP binding. It is also activated by phosphorylation at Thr-967 and Ser-1099. It is inhibited by phosphorylation at Tyr-722. It is highly expressed in brain and skeletal muscle. It is largely cytoplasmic, but can be found at the membrane and in the nucleus. Insertional mutagenesis studies in mice support a role for this protein kinase in mouse cancer oncogenesis. Elevated ROCK2 activity is linked to vascular disease, hypertension, and tumour progression (e.g., breast, gastric cancers). This description may include information annotated by UniProt and/or Google AI.