Focal adhesion kinase 1 (FAK) is a ubiquitously expressed non-receptor protein tyrosine kinase that is concentrated in the focal adhesions that form between cells growing in the presence of extracellular matrix constituents. This cellular localization is directed by a "Focal Adhesion Targeting" (FAT) sequence, a 125 amino acid sequence at the C-terminus. FAK plays an important role in migration, cell spreading, differentiation, cytoskeleton protein phosphorylation, apoptosis and acceleration of the G1 to S phase transition of the cell cycle. It associates with several different signaling proteins such as Src-family PTKs, p130Cas, Shc, Grb2, PI 3-kinase, and paxillin. This enables 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. FAK is also linked to oncogenes at biochemical and functional levels. Increased expression and/or activity of FAK in various tumors has been correlated with enhanced migration and invasiveness of human tumor cells in addition to promoting increased cell proliferation.
Human/Mouse/Rat FAK Antibody
R&D Systems | Catalog # AF4467
Key Product Details
Validated by
Knockout/Knockdown
Species Reactivity
Validated:
Human, Mouse, Rat
Cited:
Human, Rat
Applications
Validated:
Knockout Validated, Immunohistochemistry, Western Blot
Cited:
Western Blot, Immunoprecipitation, ELISA Development (Capture)
Label
Unconjugated
Antibody Source
Polyclonal Sheep IgG
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Product Specifications
Immunogen
E. coli-derived recombinant human FAK
Asp213-Thr412
Accession # Q05397
Asp213-Thr412
Accession # Q05397
Specificity
Detects human, mouse and rat FAK.
Clonality
Polyclonal
Host
Sheep
Isotype
IgG
Scientific Data Images for Human/Mouse/Rat FAK Antibody
Detection of Human, Mouse, and Rat FAK by Western Blot.
Western blot shows lysates of MCF‑7 human breast cancer cell line, HUVEC human umbilical vein endothelial cells, Neuro‑2A mouse neuroblastoma cell line, and Rat‑2 rat embryonic fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat FAK Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4467) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for FAK at approximately 125 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.FAK in Human Brain.
FAK was detected in immersion fixed paraffin-embedded sections of human brain (hippocampus) using 3 µg/mL Human/Mouse/Rat FAK Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4467) overnight at 4 °C. Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.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 1 µg/mL of Sheep Anti-Human/Mouse/Rat FAK Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4467) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). 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 (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.Detection of Human FAK by Western Blot
ADAM9 affects auto‐/paracrine MEK/ERK signaling. (A) Western blot of phosphorylation of MEK1/2 (S217/221) and ERK1/2 (T202/204) in AsPC‐1 and MiaPaCa‐2 cells following ADAM9 expression silencing. In AsPC‐1 cells, we observed decreased pFAK (Y397), which was absent in MiaPaCa‐2 cells. Quantitation of pFAK, pMEK1/2, and pERK1/2 in AsPc‐1 and MiaPaCa‐2 cells from three independent experiments. (B) Western blot analysis shows decreased phosphorylation of MEK1/2 (S217/221) and ERK1/2 (T202/204) in HUVEC cells following 30 min of incubation with cancer cell conditioned medium from both AsPC‐1 and MiaPaCa‐2 cells. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30556643), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Human/Mouse/Rat FAK Antibody
Application
Recommended Usage
Immunohistochemistry
5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain (hippocampus)
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.
Western Blot
1 µg/mL
Sample: HUVEC human umbilical vein endothelial cells, MCF-7 human breast cancer cell line, Neuro-2A mouse neuroblastoma cell line, and Rat-2 rat embryonic fibroblast cell line
Sample: HUVEC human umbilical vein endothelial cells, MCF-7 human breast cancer cell line, Neuro-2A mouse neuroblastoma cell line, and Rat-2 rat embryonic fibroblast cell line
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 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
Background: FAK
Long Name
Focal adhesion kinase 1
Alternate Names
FADK1, PTK2
Gene Symbol
PTK2
UniProt
Additional FAK Products
Product Documents for Human/Mouse/Rat FAK Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human/Mouse/Rat FAK Antibody
For research use only
Related Research Areas
Citations for Human/Mouse/Rat FAK Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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