Product Name:
CDK9-pT186
Product Number:
ab-pk575
Target Full Name: Cyclin-dependent protein-serine kinase 9
Target Alias: C-2K; CDC2L4; Cell division cycle 2-like protein kinase 4; Cyclin-dependent kinase 9; Kinase CDK9; PITALRE; TAK
Product Type Specific: CDK9 protein kinase phosphosite-specific antibody
Antibody Code: PK575
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: T186
Protein UniProt: P50750
Protein SigNET: CDK9
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human CDK9 sequence peptide Cat. No.: PE-04AJS99
Antibody Immunogen Sequence: NRY(pT)NRV(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues N183 to V189; in the protein kinase catalytic domain activation T loop region between subdomains VII and VIII. T186 phosphorylation stimulates phosphotransferase activity by promoting cyclin and substrate binding. Phosphorylation induces binding to EDG-1, RNA and TEAD1. This is one of the major in vivo phosphorylation sites in CDK9 (≥33 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). CDK9 is known to be phosphorylated at this site in vitro by CDK9.
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 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 | Zebra fish | Honey bee | Sea urchin.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 40-45 kDa.
Antibody Specificity: Very high
Antibody Cross Reactivity: No significant cross-reactive proteins detected in A431, HEK-293, and HeLa cells and in sea star oocytes, except weak ~70 and ~100 kDa proteins in sea star oocytes.
Related Product 1: CDK9 (183-189) pT186 antibody blocking peptide (Cat. No.: PE-04AJS99)
Related Product 2: CDK7/9Subtide kinase substrate peptide (Cat. No.: PE-01BGR99)
Related Product 3: RNA polymerase II-CT KinSub - heptapeptide repeat from C-terminus peptide; CDKtide protein kinase substrate peptide
Related Product 4: CDK9 S347 phosphorylation antibody (Cat. No.: AB-PK574)
Scientific Background: CDK9 is a protein-serine/threonine kinase of the CMGC group and CDK family. It is a protein kinase that is involved in chromatin binding and transcription regulator region DNA binding. It acts as the catalytic subunit of the Positive Transcription Elongation Factor b (P-TEFb) complex, which is crucial for RNA polymerase II-directed transcription elongation. CDK9 pairs with regulatory partners, primarily Cyclin T1, T2, or K, to form the active P-TEFb complex. CDK9 often accumulates on chromatin to regulate gene expression. It regulates gene expression by phosphorylating RNA polymerase II, allowing the transition from abortive to productive transcription. It phosphorylates the C-terminal domain (CTD) of RNA polymerase II, as well as negative elongation factors (DSIF and NELF), promoting the release of paused RNA polymerase II. Phosphorylation at Ser-175 increases phosphotransferase activity of CDK9. CDK9 is composed of an N-terminal lobe (amino acids 16-108) and a C-terminal lobe (residues 109-330), with a conserved ATP-binding site. It shuttles between the nucleus and cytoplasm, with nuclear localization promoted by cyclin T1. Due to its role in regulating short-lived survival proteins, CDK9 is often dysregulated in cancer cells, and may be a therapeutic target. This description may include information annotated by UniProt and/or Google AI.