Product Name:
Cas-L-pY166
Product Number:
ab-pn505
Target Full Name: Enhancer of filamentation 1
Target Alias: NEDD9; Neural precursor cell expressed developmentally down-regulated protein 9
Product Type Specific: Cas-L phosphosite-specific antibody
Antibody Code: PN505
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y166
Protein UniProt: Q14511
Protein SigNET: CasL
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human Cas-L sequence peptide Cat. No.: PE-04AFA99
Antibody Immunogen Sequence: GYV(pY)EYP(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues G163 to P169; in the region between the SH3_9 and Serine_rich domains. This is the second major in vivo phosphorylation site in Cas-L. The effect of Y166 phosphorylation is unclear. This is the second major in vivo phosphorylation site in Cas-L (≥569 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus).
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: 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 HeLa cells. 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 | Rhesus Macaque | Rat | Mouse | Platypus.
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: High
Antibody Cross Reactivity: Medium cross-reactive ~48 kDa protein detected in A431 and HeLa cells.
Related Product 1: CasL (163-169) pY166 antibody blocking peptide (Cat. No.: PE-04AFA99)
Scientific Background: CasL (Crk-associated substrate lymphocyte type; NEDD9; HEF1) is a ~105-115 kDa docking protein which plays a central coordinating role for tyrosine-kinase-based signalling related to cell adhesion. It features an SH3 domain, a substrate domain with multiple tyrosine-phosphorylation sites, and an helix-loop-helix (HLH) motif. It functions in transmitting growth control signals between focal adhesions at the cell periphery and the mitotic spindle in response to adhesion or growth factor signals initiating cell proliferation. It plays a central role in integrin-dependent signalling, cell migration, and actin cytoskeletal organization, particularly in T cells, B cells, often by acting as a docking platform for CRKL and FAK. It may also participate in integrin beta-1 or B cell antigen receptor (BCR) mediated signalling in B- and T-cells. It is essential for efficient T cell migration and infiltration into inflamed tissues. Deficiency of CasL leads to increased membrane blebbing, reduced cortical F-actin, and erratic movement. It becomes tyrosine-phosphorylated upon integrin-mediated adhesion or T cell receptor (TCR) stimulation. It acts as a scaffold promoting adhesion, migration, and invasion, and is frequently upregulated in various cancers. This description may include information annotated by UniProt and/or Google AI.