The experiment involved the Jurkat cell line and included treatment with hydrogen peroxide at a concentration of 100 uM for 10 minutes. Serum starvation was also conducted. Following treatment, the cells were fixed for 20 minutes on ice and permeabilized using 0.2% Triton X. The primary antibody was diluted to a concentration of 1ug/mL, and the secondary antibody used was AP182F at a dilution of 1:1000.
ABT135
Anti-phospho-FAK (Tyr397) Antibody
from rabbit, purified by affinity chromatography
Synonym(s):
Focal adhesion kinase 1, FADK 1, Protein-tyrosine kinase 2, pp125FAK
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100 μL
$794.00
About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Recommended Products
biological source
rabbit
Quality Level
antibody form
affinity isolated antibody
antibody product type
primary antibodies
clone
polyclonal
purified by
affinity chromatography
species reactivity
rat
species reactivity (predicted by homology)
human (based on 100% sequence homology), mouse (based on 100% sequence homology), Xenopus (based on 100% sequence homology), chicken (based on 100% sequence homology)
technique(s)
cell based assay: suitable
western blot: suitable
NCBI accession no.
UniProt accession no.
shipped in
wet ice
target post-translational modification
phosphorylation (pTyr397)
Gene Information
human ... PTK2(5747)
General description
Focal adhesion kinase (FAK) is a non receptor protein tyrosine kinase discovered as a substrate for Src and a key element in integrin signaling. FAK plays a central role in cell spreading, differentiation, migration, cell death and acceleration of the G1 to S phase transition of the cell cycle. FAK regulation includes phosphorylation at multiple tyrosine and serine residues. Phosphorylation of tyrosine is generally associated with positive regulation and growth promotion; however, dephosphorylation at these sites occurs as cells enter mitosis (M-Phase). In contrast, serine phosphorylation either remains high or is increased as cells enter mitosis and may play a role in focal adhesion disassembly. FAK and its phosphorylation states have been implicated in cancer metastasis, tumor cell survival, and adhesion-independent growth. Recent evidence indicates that elevation of FAK activity in human carcinoma cells is associated with increased invasive potential. A central role in tumor formation and progression suggests that FAK is an attractive target for therapeutic intervention. Activation of FAK by integrin clustering leads to autophosphorylation at Tyr397.
Specificity
This antibody recognizes FAK phosphorylated at Tyr397.
Immunogen
Epitope: Phosphorylated Tyr397
KLH-conjugated linear peptide corresponding to human FAK phosphorylated at Tyr397.
Application
Detect phospho-FAK (Tyr397) using this Anti-phospho-FAK (Tyr397) Antibody validated for use in WB, Cell Function Assay.
Peptide Inhibition Assay Analysis: A 1:1,000 dilution from a representative lot peptide blocked on RAT2 cells transfected with Src.
Research Category
Cell Structure
Cell Structure
Research Sub Category
Adhesion (CAMs)
Adhesion (CAMs)
Quality
Evaluated by Western Blot in untransfected and Src-transfected RAT2 cell lysates.
Western Blot Analysis: A 1:1,000 dilution of this antibody detected phospho-FAK (Tyr397) in 10 µg of untransfected and Src-transfected RAT2 cell lysates.
Western Blot Analysis: A 1:1,000 dilution of this antibody detected phospho-FAK (Tyr397) in 10 µg of untransfected and Src-transfected RAT2 cell lysates.
Target description
~125 kDa observed.
An uncharacterized band appears at ~190 kDa in some lysates.
An uncharacterized band appears at ~190 kDa in some lysates.
Linkage
Replaces: 04-974
Physical form
Affinity purified
Purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Storage and Stability
Stable for 1 year at 2-8°C from date of receipt.
Analysis Note
Control
untransfected and Src-transfected RAT2 cell lysates
untransfected and Src-transfected RAT2 cell lysates
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Where can I find the protocol for the experiment shown in the histogram of product ABT135, specifically for the hydrogen peroxide treatment details?
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