Product Name: Syk-pY323
Product Number: AB-PK821
Size: 25 µg      Price:89.00
      $US
Target Full Name: Spleen protein-tyrosine kinase

Target Alias: DKFZp313N1010; FLJ25043; FLJ37489; Kinase Syk; KSyk; Spleen tyrosine kinase; Syk; CCDS6688.1; ENSG00000165025

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK821

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: Y323

Protein UniProt: P43405

Protein SigNET: P43405

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human Syk sequence peptide Cat. No.: PE-04ALE99

Antibody Immunogen Sequence: FNP(pY)EPE(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues F320 to E326; In the region between the SH2 and kinase catalytic domains. The major site of phosphorylation in Syk.

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: 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 Jurkat cells. 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 67-77 kDa.

Antibody Specificity: Medium-High

Antibody Cross Reactivity: Almost no significant cross-reactivities detected in T98G cells and sea star oocytes, but with SDS-PAGE sample buffer extraction in Jurkat and A431 cells, get detection of cross-reactive proteins from 120-140 kDa.

Related Product 1: Syk-pY323 blocking peptide

Related Product 2: Syk-pY525+pY526 phosphosite-specific antibody (Cat. No.: AB-PK823)

Related Product 3: SykSubtide - Syk protein kinase substrate peptide

Scientific Background: Syk is one of several protein kinases that can phosphorylate KinSub4DDDYV. Syk is a protein-tyrosine kinase that is a member of the TK group of protein kinases in the Syk family. It is moderate to highly expressed in most tested human tissues, particularly in hematopoietic cells. It mediates cell proliferation, differentiation, and phagocytosis. In B cells, Syk plays a crucial role in intracellular signalling induced by oxidative stress as well as antigen receptor engagement. Syk is activated by phosphorylation at Y525 and Y526. Phosphorylation of Y348 and Y352 enhances the binding, phosphorylation and activation of PLC-gamma and the early phase of Ca(2+) mobilization via a PI 3-kinase-independent pathway. Phosphorylation of Y131 of Syk inhibits interaction with Bcr. Phosphorylation of Y323 of Syk inhibits B cell Ag receptor signalling and strongly dampens the Ca(2+) signal. Syk appears to be an oncoprotein (OP) and a tumour suppressor protein (TSP). Syk is an oncoprotein in retinoblastoma, which is upregulated and is required for tumour cell survival. Syk is a potential tumour suppressor in breast cancer, where it absence in primary breast tumours is correlated with poor outcomes. Syk deficient cells have increased motility that is restored to normalcy by replacement with wild-type Syk. It has been linked with the development of allergy, asthma, inflammation, gastric cancer, acute lymphoblastic leukemia, and acute myeloid leukemia.
Figure 1. Epitope mapping of Syk-pY323 antibody with similar phosphopeptides on dot blots.


Figure 2. Identification of phosphosites related to Syk-pY323.