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Product Specifications

Analytical sensitivity

<0.9 U/mL

Assay range

1.6-100 U/mL

Sample type/volume

Cell Lysate
10 µL

Hands-on time

1 hr 20 min

Time-to-result

4 hr

Homogenous (no wash)

No

Interassay CV

8%

Intraassay CV

4%

Instrument

Colorimetric Microplate Reader

Product size

96 Tests

Contents

Pre-coated 96 well plate
Standard
Standard Dilution Buffer
Detection Antibody
anti-rabbit-HRP
HRP Diluent
Wash Buffer
Chromogen
Stop Solution
Adhesive Plate Covers

Shipping conditions

Wet ice

Storage

2-8°C

Protein name

FAK

Target site-specificity

Phosphorylated Tyr397

Protein subtype

Cytoplasmic Tyrosine Kinases

Protein family

Kinases & Inhibitors

Species (tested)

Human, Mouse

Assay kit format

Sandwich ELISA Kit

Detector antibody conjugate

HRP

Label or dye

HRP

About This Kit

The Human FAK (Phospho) [pY397] ELISA Kit is a solid-phase sandwich Enzyme-Linked Immunosorbent Assay (ELISA) designed to detect and quantify the level of FAK (Phospho) [pY397]) in fresh or frozen human and mouse cell lysates. The assay recognizes both natural and recombinant FAK (Phospho) [pY397].

Principle of the method

A monoclonal capture antibody specific for FAK (Phospho) [pY397] has been coated onto the wells of the 96-well plate. During the first incubation, standards of known content and unknown samples are pipetted into the wells and the antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for the target protein is added to the wells and serves as a detection antibody by binding to the immobilized protein captured during the first incubation. After washing, a horseradish peroxidase labeled anti-rabbit IgG is added. This binds to the detection antibody to complete the four member sandwich. After a third incubation and washing to remove all the unbound enzyme, a substrate solution (TMB) is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of target protein present in the original specimen and the optical density can be read on a standard microplate reader.

Rigorous validation

Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Target information

Focal Adhesion Kinase (FAK) is a 125 kDa non-receptor protein tyrosine kinase that acts as a substrate for Src and is a key element of integrin signaling. FAK plays an important role in cell spreading, differentiation, migration, cell death, and acceleration of the G1 to S phase transition of the cell cycle. FAK has a central catalytic domain and a C-terminal tail that localizes it to focal adhesions, which are sites where cells attach to the extracellular matrix via surface integrin receptors. Increased FAK tyrosine phosphorylation occurs upon integrin engagement with fibronectin. Adhesion of murine NIH3T3 fibroblasts to fibronectin promotes association of the Grb2 adapter protein and c-Src PTK with FAK in vivo, and also results in activation of the ERK2 MAP kinase. In v-Src-transformed NIH3T3, the association of v-Src, Grb2, and Sos with FAK is independent of cell adhesion to fibronectin. In vitro the Grb2 SH2 domain binds directly to tyrosine-phosphorylated FAK, and the binding site has been identified as Tyr925 by site directed mutagenesis. Several transcript variants encoding different isoforms have been found for the FAK gene, but the full-length natures of only three of them have been determined.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Gene aliases : FADK, FAK, FAK1, FRNK, Kiaa4203, mKIAA4203, p125FAK, pp125FAK, PPP1R71, PTK2

Gene ID : (Human) 5747, (Mouse) 14083

Gene symbol : PTK2, Ptk2

Protein Aliases : FADK 1, FAK-related non-kinase polypeptide, Focal adhesion kinase 1, focal adhesion kinase isoform FAK-Del33, Focal adhesion kinase-related nonkinase, focal ashension kinase 1, FRNK, p125FAK, pp125FAK, PPP1R71, Protein phosphatase 1 regulatory subunit 71, protein phosphatase 1, regulatory subunit 71, Protein-tyrosine kinase 2, PTK2 protein tyrosine kinase 2

UniProt ID (Human) Q05397, (Mouse) Q5DTH7

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