Product Name: Nek2-pT170+pS171
Product Number: AB-PK733
Size: 25 µg      Price:89.00
      $US
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: Protein kinase phosphosite-specific antibody

Antibody Code: PK733

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: T170+S171

Protein UniProt: P51955

Protein SigNET: P51955

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human Nek2 sequence peptide Cat. No.: PE-04ABO90

Antibody Immunogen Sequence: HD(pT)(pS)FAK(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues H168 to K174; In protein kinase catalytic domain activation T-loop between subdomains VII and VIII.

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: 0.3 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 Species Reactivity: Human

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 47-55 kDa.

Related Product 1: Nek2-pT170+pS171 blocking peptide

Related Product 2: Nek2-pS171 phosphosite-specific antibody (Cat. No.: AB-PK732)

Scientific Background: Nek2 (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 can phosphorylate CROCC, CEP250 and NINL, ultimately removing their ability to form a complex in the centrosomes. The proteins mitotic checkpoint proteins CDC20 and MAD2L1 are also phosphorylated and mediated by Nek2. The effect on chromosome condensation is via HMGA2. 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. Nek2 dysfunction has been associated with both primary and secondary breast tumours.
Figure 1. Epitope mapping of Nek2-pT170+pS171 antibody with similar phosphopeptides on dot blots.


Figure 2. Identification of phosphosites related to Nek2-pT170+pS171.