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Invitrogen™ Human FAK (Phospho) [pY397] ELISA Kit

Catalog No. KHO0441
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KHO0441 96 Tests
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ELISA

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,...

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.
TRUSTED_SUSTAINABILITY

Specifications

Accession Number P34152, Q05397
Assay Range 1.6-100 U/mL
Assay Sensitivity <0.9 U/mL
Concentration 1.6 U/mL
Content And Storage Pre-coated plate(s), standard or calibrator, detector antibody, HRP conjugate, diluents, wash buffer, substrate(s), and stop solution. Store kit at 2-8°C. See product manual for detailed contents and storage conditions for maximum stability.
Conjugate HRP
Immunoassay Kit Format Sandwich ELISA Kit
Product Type ELISA
Protein Family Kinases & Inhibitors
Sample Type Cell Lysates
Substrate Type Cytoplasmic Tyrosine Kinases
Target Species Human, Mouse
For Use With (Equipment) Absorbance Microplate Reader
Gene Alias CG10023; CG10023-PA; CG10023-PB; CG10023-PC; CG10023-PD; CG10023-PF; CG10023-PG; CT28129; DFAK; DFak56; Dmel\CG10023; Dmel_CG10023; DmFAK; EC 2.7.10.2; Fadk; FADK 1; Fak; Fak1; Fak56; fak56D; FAK65D; Fak-PA; Fak-PB; Fak-PC; Fak-PD; Fak-PF; Fak-PG; FAK-related non-kinase polypeptide; focal adhesion kinase; Focal adhesion kinase 1; focal adhesion kinase homolog; focal adhesion kinase isoform FAK-Del33; focal adhesion kinase pp125FAK; focal adhesion kinase-related nonkinase; focal ashension kinase 1; FRNK; I79_019430; Kiaa4203; mKIAA4203; p125FAK; p41/p43FRNK; pFAK; pp125 PTK2; pp125FAK; PPP1R71; Protein phosphatase 1 regulatory subunit 71; protein phosphatase 1, regulatory subunit 71; protein tyrosine kinase 2; protein tyrosine kinase 2 L homeolog; protein-tyrosine kinase; protein-tyrosine kinase 2; PTK2; PTK2 protein tyrosine kinase 2; ptk2.L; tyrosine kinase; XELAEV_18032408mg; XPFAKCG10023; CG10023-PA; CG10023-PB; CG10023-PC; CG10023-PD; CG10023-PF; CG10023-PG; CT28129; DFAK; DFak56; Dmel\CG10023; Dmel_CG10023; DmFAK; EC 2.7.10.2; Fadk; FADK 1; Fak; Fak1; Fak56; fak56D; FAK65D; Fak-PA; Fak-PB; Fak-PC; Fak-PD; Fak-PF; Fak-PG; FAK-related non-kinase polypeptide; focal adhesion kinase; Focal adhesion kinase 1; focal adhesion kinase homolog; focal adhesion kinase isoform FAK-Del33; focal adhesion kinase pp125FAK; focal adhesion kinase-related nonkinase; focal ashension kinase 1; FRNK; I79_019430; Kiaa4203; mKIAA4203; p125FAK; p41/p43FRNK; pFAK; pp125 PTK2; pp125FAK; PPP1R71; Protein phosphatase 1 regulatory subunit 71; protein phosphatase 1, regulatory subunit 71; protein tyrosine kinase 2; protein tyrosine kinase 2 L homeolog; protein-tyrosine kinase; protein-tyrosine kinase 2; PTK2; PTK2 protein tyrosine kinase 2; ptk2.L; tyrosine kinase; XELAEV_18032408mg; XPFAK;
Gene ID (Entrez) 14083, 5747
Gene Symbol PTK2
Incubation Time 4 hr.
Interassay CV 8%
Intraassay CV 4%
Kit 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, Standard, Standard Dilution Buffer, Detection Antibody, anti-rabbit-HRP, HRP Diluent, Wash Buffer, Chromogen, Stop Solution, Adhesive Plate Covers
Label or Dye HRP
Marker Gene FAK
Quantity 96 Tests
Regulatory Status RUO
Sample Volume Cell Lysate 10 μL
Shipping Condition Wet Ice
Storage Requirements 2-8°C
Target FAK
Test Time 1 hr 20 min
Total Assay Time 4 hr
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