Product Name:
Nek2-pS171
Product Number:
ab-pk732
Target Full Name: NIMA (never-in-mitosis)-related protein-serine kinase 2
Target Alias: HSPK 21; Kinase Nek2; Nek2A; NIMA (never in mitosis gene a)-related kinase 2; NimA-like protein kinase 1; NimA-related protein kinase 2; NLK1
Product Type Specific: Nek2 protein kinase phosphosite-specific antibody
Antibody Code: PK732
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S171
Protein UniProt: P51955
Protein SigNET: Nek2
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human Nek2 sequence peptide Cat. No.: PE-04ABP99
Antibody Immunogen Sequence: HDT(pS)FAK(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues H168 to K174; in the protein kinase catalytic domain activation T loop region between subdomains VII and VIII. S171 phosphorylation stimulates phosphotransferase activity. This is one of the major in vivo phosphorylation sites in Nek2 (≥5 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). Nek2 is known to be phosphorylated at this site in vitro by NEK2.
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: 0.75 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 | Chimpanzee | Rhesus Macaque | Dog | Mouse | Platypus | Chicken | Frog | Sea urchin.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 47-55 kDa.
Antibody Specificity: High-very high
Related Product 1: Nek2 (168-174) pS171 antibody blocking peptide (Cat. No.: PE-04ABP99)
Related Product 2: Nek2 T170+S171 phosphorylation antibody (Cat. No.: AB-PK733)
Related Product 3: Nek6 S206 phosphorylation antibody (Cat. No.: AB-PK734)
Related Product 4: Nek7 T191+S195 phosphorylation antibody (Cat. No.: AB-PK735)
Related Product 5: Nek1 S1052 phosphorylation antibody (Cat. No.: AB-PK940)
Scientific Background: Nek2 (NIMA-related kinase 2; NIMA) is a protein-serine/threonine kinase of the Other group and Nek family. It regulates bipolar spindle formation and centrosome separation in mitotic cells, and affects chromatin condensation in meiotic cells. NEK2 is crucial for centrosome disjunction (splitting) and bipolar spindle formation, ensuring proper chromosome segregation. It is highly expressed in the S and G2 phases, with low levels in G1 phase, and peaking just before mitosis. It.localizes to the centrosome, where it phosphorylates proteins like C-Nap1 to trigger centrosome splitting. Nek2 can also phosphorylate CROCC and NINL, ultimately removing their ability to form a complex in the centrosomes. The mitotic checkpoint proteins CDC20 and MAD2L1 are also phosphorylated Nek2. Its effect on chromosome condensation is mediated via HMGA2. There are two main splice variants, Nek2A is the longer variant, and Nek2B, which is more stable in early mitosis. The N-terminal region features the kinase catalytic domain, whereas the C-terminal domain contains a leucine zipper (LZ) motif for dimerization and a PP1-binding site (specifically in Nek2A). The ability for Nek2 to activate Nek11 is lost with a K37R mutation. Nek2 kinase phosphotransferase activity can be increased two-fold with a T170E, or S171D, or T175E mutation, while the phosphotransferase activity can be decreased two-fold with a T175A mutation. Complete kinase phosphotransferase activity can be lost with a T179A, T179E, S241A, or S241D mutations. Nek2 is linked to Plexiform Neurofibromas, and chromosome polyploidy. Polyploidy is linked to breast cancer and melanoma, and most often affects smooth muscle, liver, and breast tissues. Upregulation of Nek2 has been associated with both primary and secondary breast tumours and prostate cancer, contributes to resistance to chemotherapy drugs. This description may include information annotated by UniProt and/or Google AI.