Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Knockout Validated, Immunohistochemistry, Western Blot, Simple Western

Cited:

Western Blot, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 495919
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Product Specifications

Immunogen

E. coli-derived recombinant human FAK
Asp213-Thr412
Accession # Q05397

Specificity

Detects human FAK in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human FAK Antibody

Detection of Human FAK antibody by Western Blot.

Detection of Human FAK by Western Blot.

Western blot shows lysates of MCF-7 human breast cancer cell line, A549 human lung carcinoma cell line, Huh-7 human hepatoma cell line, and HUVEC human umbilical vein endothelial cells. PVDF membrane was probed with 2 µg/mL of Human FAK Monoclonal Antibody (Catalog # MAB4467) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF007). A specific band was detected for FAK at approximately 125 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.
FAK antibody in Human Brain by Immunohistochemistry (IHC-P).

FAK in Human Brain.

FAK was detected in immersion fixed paraffin-embedded sections of human brain (hippocampus) using Human FAK Monoclonal Antibody (Catalog # MAB4467) at 25 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Western Blot Shows Human FAK Antibody Specificity by Using Knockout Cell Line.

Western Blot Shows Human FAK Specificity by Using Knockout Cell Line.

Western blot shows lysates of HEK293T human embryonic kidney parental cell line and FAK knockout HEK293T cell line (KO). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human FAK Monoclonal Antibody (Catalog # MAB4467) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for FAK at approximately 135 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human FAK by Simple WesternTM.

Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line and A549 human lung carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for FAK at approximately 120 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human FAK Monoclonal Antibody (Catalog # MAB4467). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Human FAK by Western Blot

Detection of Human FAK by Western Blot

Phosphorylation of FAK following treatment of AF cells derived from non-degenerate IVDs with 1.0 Hz CTS, with and without pre-treatment with RGD or RAD peptides.AF cells derived from non-degenerate IVDs (n = 4) were treated+/− RGD (50 µg/ml) or RAD (50 µg/ml) peptides, and mechanically stimulated (10% CTS, 1.0 Hz frequency) in serum-free media and total protein extracted at timepoints of 5 and 20 minutes. Mechanically stimulated and unstimulated,+/− RGD or RAD peptides, non-degenerate protein samples (5 µg/well) exposed to A) 5 minutes and B) 20 minutes of CTS, were separated using 10% SDS-PAGE and probed using primary antibodies against phosphorylated FAK. Blots were then stripped using a stripping buffer, re-blocked and probed using an antibody against total FAK protein. C) The density of bands were quantified using a Syngene imaging system and the ratio of phosphorylated: total FAK protein normalised to timepoint controls and plotted as % change. *denotes a significant change (p≤0.05) between treatment groups. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24039840), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human FAK Antibody

Application
Recommended Usage

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain (hippocampus)

Knockout Validated

FAK is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in FAK knockout HEK293T cell line.

Simple Western

20 µg/mL
Sample: MCF-7 human breast cancer cell line and A549 human lung carcinoma cell line,

Western Blot

2 µg/mL
Sample: MCF-7 human breast cancer cell line, A549 human lung carcinoma cell line, Huh-7 human hepatoma cell line, and HUVEC human umbilical vein endothelial cells

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: FAK

Focal adhesion kinase 1 (FAK), also known as FAK1 and PTK2, is a ubiquitously expressed non-receptor protein tyrosine kinase that is concentrated in focal adhesions. This cellular localization is directed by a C-terminal 125 amino acid "Focal Adhesion Targeting" (FAT) sequence. FAK plays an important role in migration, cell spreading, differentiation and apoptosis. It associates with several different signaling proteins, such as Src-family PTKs, p130Cas, Shc, Grb2, PI 3-kinase, and Paxillin. These associations enable FAK to function within a network of integrin-stimulated signaling pathways, leading to the activation of targets such as the ERK and JNK mitogen-activated protein kinase pathways. Increased expression and/or activity of FAK in various cancers has been correlated with enhanced proliferation, migration and invasiveness of human tumor cells.

Long Name

Focal adhesion kinase 1

Alternate Names

FADK1, PTK2

Entrez Gene IDs

5747 (Human); 14083 (Mouse); 25614 (Rat)

Gene Symbol

PTK2

UniProt

Additional FAK Products

Product Documents for Human FAK Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human FAK Antibody

For research use only

Citations for Human FAK Antibody

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs

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Associated Pathways

VEGF - VEGF R2 Signaling Pathways VEGF - VEGF R2 Signaling Pathway Thumbnail