Product Name:
GIT1-pY545
Product Number:
ab-pn517
Target Full Name: ARF GTPase-activating protein GIT1
Target Alias: ARF GAP GIT1; ARF GTPase-activating protein GIT1; CAT-1; CAT1; Cool-associated and tyrosine-phosphorylated protein 1; G protein-coupled receptor kinase interacting ArfGAP 1; G protein-coupled receptor kinase interactor 1; G protein-coupled receptor kinase-interactor 1; GIT1; GRK-interacting protein 1
Product Type Specific: GIT1 phosphosite-specific antibody
Antibody Code: PN517
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y545
Protein UniProt: Q9Y2X7
Protein SigNET: GIT1
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human GIT1 sequence peptide Cat. No.: PE-04AEN99
Antibody Immunogen Sequence: DAI(pY)SVH(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues D542 to H548; in the last third of the protein before the GIT1_C domain. Y545 phosphorylation inhibits interaction with His-5 and PXN and induces cell motility. This is the major in vivo phosphorylation site in GIT1 (≥767 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 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 | Chimpanzee | Rhesus Macaque | Dog | Rat | Mouse.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 85-95 kDa.
Antibody Specificity: Very high
Antibody Cross Reactivity: No significant cross-reactive proteins detected in A431 and Jurkat cells.
Related Product 1: GIT1 (542-548) pY545 antibody blocking peptide (Cat. No.: PE-04AEN99)
Related Product 2: GIT1 expression antibody (Cat. No.: AB-NN261-1)
Scientific Background: GIT1 is a GTPase-activating protein for the ADP ribosylation factor family. It may serve as a multi-functional scaffold to bring together molecules to form signalling modules controlling vesicle trafficking, adhesion and cytoskeletal reorganization. It increases the speed of cell migration, as well as the size and rate of formation of protrusions, possibly by targeting PAK1 to adhesions and the leading edge of lamellipodia. It sequesters inactive non-tyrosine-phosphorylated paxillin in cytoplasmic complexes. It also acts as a scaffold for PIX (PAK-interacting exchange factor), and phospholipase C-gamma (PLCg) to regulate cell spreading, migration, and membrane protrusion. It features an N-terminal ARF-GAP domain, three ankyrin repeats, a Spa2-homology domain (SHD), and a carboxyl-terminal paxillin-binding site. It is found in focal adhesions, the cytoplasm, and the cytosol. GIT1 acts in focal adhesions, where it regulates membrane trafficking between the plasma membrane and endosomes. It plays a role in synaptic transmission and has been implicated in ADHD-like behaviors in animal models. GIT1 is also involved in angiogenesis and vascular permeability. Overexpression of GIT1 is associated with tumour progression and metastasis in various cancers (e.g., liver and lung).This description may include information annotated by UniProt and/or Google AI.