Product Name:
LIMK1-pT508
Product Number:
ab-pk681
Target Full Name: LIM domain kinase 1
Target Alias: Kinase LIMK1; KIZ; KIZ-1; LIMK; LIMK-1; CCDS5563.1; P53667; Q75MU0; Q75MU4; ENSG00000106683
Product Type Specific: LIMK1 protein kinase phosphosite-specific antibody
Antibody Code: PK681
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: T508
Protein UniProt: P53667
Protein SigNET: LIMK1
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human LIMK1 sequence peptide Cat. No.: PE-04ANI99
Antibody Immunogen Sequence: KRY(pT)VVG(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues K505 to G511; in the protein kinase catalytic domain. T508 phosphorylation stimulates phosphotransferase activity and induced interaction with NISCH. This phosphosite is located in the kinase activation loop between catalytic subdomains VII and VIII. This is one of the minor in vivo phosphorylation sites in LIMK1 (≥2 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). LIMK1 is known to be phosphorylated at this site in vitro by MRCKa (CDC42BPA, PK428), PAK1, PAK4, PAK6, and ROCK1.
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.
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: Strong immunoreactivity of a target-sized protein by Western blotting in A549 and MCF7 cells, andMedium detection in Jurkat and HepG2 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 | Dog | Rat | Mouse | Platypus | Chicken | Frog | Zebra fish | Fruit fly | Honey bee.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 72-78 kDa.
Antibody Specificity: Medium- very high
Antibody Cross Reactivity: No significant cross-reactive proteins detected in A549 cells and very little in HepG2 and MCF7 cells. In HEK-293 cells, phenylarsine oxide (PAO) decreased 100 + 80 kDa proteins; in Jurkat cells, PAO increased 50 + 60 kDa proteins, and PAO decreased 110 + 95 kDa proteins. In MCF7 cells, insulin increased detection of a 75 kDa protein.In sea star oocytes undergoing meiotic maturation, at least four strong cross-reactive proteins of ~90, ~140, ~200 and ~250 kDa were evident.
Related Product 1: LIMK1 (505-511) pT508 antibody blocking peptide (Cat. No.: PE-04ANI99)
Scientific Background: LIMK1 is a protein-serine/threonine kinase of the TKL group and LISK family, and it is involved in regulation of cell motility, cell morphology, cytokinesis and cell cycle progression. It is mainly found in the cytoplasm and is often associated with microtubules and actin at focal adhesions, stress fibers, and lamellipodia. It acts as a key regulator of the actin cytoskeleton, by phosphorylating and inactivating cofilin, an actin-depolymerizing factor. Cofilin normally acts to depolymerize actin filaments (F-actin). It is primarily expressed in the nervous system, and LIMK1 controls actin filament dynamics, influencing cell structure, migration, and neuronal growth. ROCK1, PAK1 and PAK4 act upstream of LIMK1, and it is activated via phosphorylation of an active-site threonine. Phosphorylation at Ser-323 and Thr-508 increases phosphotransferase activity. Activation of LIMK1 by ROCK1 can be fully inhibited with a T508A mutation. LIMK1 features two N-terminal LIM domains (zinc fingers for protein-protein interactions) and a C-terminal protein kinase domain. LIMK1 will directly act on CFL1 (cofilin-1), CFL2 (cofilin-2), and DSTN (destrin), preventing the ability of these cytoskeletal proteins to polymerize. It also catalyzes an inactivating phosphorylation on TPPP to inhibit depolymerization and destablilization of actin and tubulin. Overexpression leads to developing invasive phenotype and metastasis of cancers. Cell cycle progression, motility, and differentiation are all affected by this inhibited actin polymerization. In LIMK1, the mutations C84S, G177E + L178A, and T508E enhance aggregation of actin whereas D460N, and R503G + K504A + K505A fully inhibits actin aggregation, and T508V/E partially inhibits actin aggregation. LMIK1 is essential for neural function, with high expression in the brain. Hemizygosity of the LIMK1 gene is linked to the impaired visuospatial cognitive abilities seen in Williams syndrome. LIMK1 is associated with cancer cell invasion and migration, particularly in prostate cancer, where high levels can promote tumour progression. This description may include information annotated by UniProt and/or Google AI.