Product Name: HGS-pY216
Product Number: AB-PN519
Size: 25 µg      Price:89.00
      $US
Target Full Name: Hepatocyte growth factor-regulated tyrosine kinase substrate

Target Alias: Human growth factor-regulated tyrosine kinase substrate; Protein pp110; Vps27; ZFYVE8

Product Type Specific: HGS phosphosite-specific antibody

Antibody Code: PN519

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: Y216

Protein UniProt: O14964

Protein SigNET: O14964

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human HGS (Hrs) sequence peptide Cat. No.: PE-04AFB95
Antibody Immunogen Sequence: EPC(pY)EQL(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues E213 to L219; In the FYVE domain. This is the major in vivo phosphorylation site in HGS.

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: 0.2 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 100-110 kDa.

Antibody Specificity: Very 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.

Related Product 1: HGS-pY216 blocking peptide

Scientific Background: HGS mediates intracellular signal transduction from cytokines and growth factors. When associated with STAM, it suppresses DNA signalling upon stimulation by IL-2 and GM-CSF. It has been implicated as a direct effector of PI3-kinase in vesicular pathway via early endosomes, regulating trafficking to early and late endosomes by recruiting clathrin, concentrating ubiquitinated receptors within clathrin-coated regions, and down-regulating receptor-tyrosine kinases via multivesicular body (MVBs) when complexed with STAM (ESCRT-0 complex). HGS may contribute to the efficient recruitment of SMADs to the activin receptor complex. Involved in receptor recycling via its association with the CART complex.
Figure 1. Epitope mapping of HGS-pY216 antibody with similar phosphopeptides on dot blots.


Figure 2. Identification of phosphosites related to HGS-pY216.