Revision 5

#71433Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, W-S, IP, IHC-Bond, IHC-P, IF-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

125

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q05397

Entrez-Gene Id:

5747

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
IHC Leica Bond 1:150 - 1:600
Immunohistochemistry (Paraffin) 1:300 - 1:1200
Immunofluorescence (Immunocytochemistry) 1:50 - 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier-free (BSA and azide free) version of this product see product #60557.

Specificity / Sensitivity

FAK (D5O7U) XP® Rabbit mAb recognizes endogenous levels of total FAK protein. Immunofluorescence was performed on cells plated on tissue culture grade plastic. We did not detect FAK signal on uncoated glass slides.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val793 of human FAK protein.

Background

Focal adhesion kinase (FAK) is a widely expressed cytoplasmic protein tyrosine kinase involved in integrin-mediated signal transduction. It plays an important role in the control of several biological processes, including cell spreading, migration, and survival (1). Activation of FAK by integrin clustering leads to autophosphorylation at Tyr397, which is a binding site for the Src family kinases PI3K and PLCγ (2-5). Recruitment of Src family kinases results in the phosphorylation of Tyr407, Tyr576, and Tyr577 in the catalytic domain, and Tyr871 and Tyr925 in the carboxy-terminal region of FAK (6,7).

  1. Parsons, J.T. et al. (2000) Oncogene 19, 5606-13.
  2. Schaller, M.D. et al. (1994) Mol Cell Biol 14, 1680-8.
  3. Cobb, B.S. et al. (1994) Mol Cell Biol 14, 147-55.
  4. Chen, H.C. et al. (1996) J Biol Chem 271, 26329-34.
  5. Zhang, X. et al. (1999) Proc Natl Acad Sci U S A 96, 9021-6.
  6. Calalb, M.B. et al. (1995) Mol Cell Biol 15, 954-63.
  7. Schlaepfer, D.D. et al. (1994) Nature 372, 786-791.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
CST is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 5
#71433

FAK (D5O7U) XP® Rabbit mAb

Western Blotting Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using FAK (D5O7U) XP® Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Western Blotting Image 2: FAK (D5O7U) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from control HEK293 cells (lane 1), or FAK knockout HEK293 cells (lane 2) using FAK (D5O7U) XP® Rabbit mAb, #71433, (upper) or #8457 β-Actin (D6A8) Rabbit mAb (lower). The absence of signal in FAK-knockout HEK293 cells confirms specificity of the antibody for FAK.
Western Blotting Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
Simple Western™ analysis of lysates (0.1 mg/mL) from A431 cells treated with hEGF (100 ng/mL, 30”) using FAK (D5O7U) XP® Rabbit mAb #71433. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66 - 440 kDa separation module.
Immunoprecipitation Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
Immunoprecipitation of FAK from RD cell lysate. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is FAK (D5O7U) XP® Rabbit mAb. Western blot analysis was performed using FAK (D5O7U) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.
Immunohistochemistry Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using FAK (D5O7U) XP® Rabbit mAb performed on the Leica® BOND Rx. 
Immunohistochemistry Image 2: FAK (D5O7U) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using FAK (D5O7U) XP® Rabbit mAb performed on the Leica® BOND Rx. 
Immunohistochemistry Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using FAK (D5O7U) XP® Rabbit mAb.
Immunohistochemistry Image 2: FAK (D5O7U) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using FAK (D5O7U) XP® Rabbit mAb.
Immunohistochemistry Image 3: FAK (D5O7U) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using FAK (D5O7U) XP® Rabbit mAb.
Immunohistochemistry Image 4: FAK (D5O7U) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using FAK (D5O7U) XP® Rabbit mAb.
Immunohistochemistry Image 5: FAK (D5O7U) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded HT-29 cell pellet (left, positive) and RPMI 8226 cell pellet (right, negative) using FAK (D5O7U) XP® Rabbit mAb.
Immunofluorescence Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of OVCAR3 cells (left, high-expressing) and SK-OV-3 cells (right, moderate-expressing) using FAK (D5O7U) XP® Rabbit mAb (green). Red = Propidium Iodide (PI)/RNase Staining Solution #4087.