Product Name: NLK-pT298
Product Number: AB-PK736
Size: 25 µg      Price:89.00
      $US
Target Full Name: Serine/threonine protein kinase NLK

Target Alias: Kinase NLK; LAK1; Nemo-like kinase

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK736

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: T298

Protein UniProt: Q9UBE8

Protein SigNET: Q9UBE8

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human NLK sequence peptide Cat. No.: PE-04AOF99

Antibody Immunogen Sequence: RHM(pT)QEV(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues R295 to V301; 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: 0.25 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: Human

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 64-68 kDa.
Antibody Specificity: High

Antibody Cross Reactivity: No significant cross-reactive proteins detected in phenylarsine oxide (PAO)+vanadate-treated HeLa cells, EGF-treated A431 cells and insulin-treated MCF7 cells, when these cells were homogenized in SDS-PAGE sample buffer, although a strong background >75 kDa was observed.

Related Product 1: NLK-pT298 blocking peptide

Scientific Background: NLK is a protein-serine/threonine kinase that is a member of the CMGC group of protein kinases in the MAPK family, and nmo subfamily. This kinase is highly expressed and widely distributed in most tested human tissues. NLK is activated by phosphorylation at tyrosine and threonine, and by by activin. NLK plays a role in cell fate determination and is required for the differentiation of bone marrow stromal cells. NLK also suppresses a wide range of transcription factors such as CREB by phosphorylation of the co-activator CREBBP. NLK expression increases the number of apoptotic cells but does not induce obvious cell cycle arrest. NLK suppresses Notch signalling by interfering with formation of the Notch active transcriptional complex. NLK has been linked with the development of glioblastoma multiforme (GM). Overexpression of NLK is associated with suppressed human colon cancer cell line growth. NLK inhibits human adenosquamous carcinoma cells proliferation and block cell cycle progression to S phase.
Figure 1. Epitope mapping of NLK-pT298 antibody with similar phosphopeptides on dot blots.