Recombinant
RabMAb

Anti-FAK antibody [EP695Y] (ab40794)

Overview

  • Product name
    Anti-FAK antibody [EP695Y]
    See all FAK primary antibodies
  • Description
    Rabbit monoclonal [EP695Y] to FAK
  • Specificity
    ab40794 recognises Focal adhesion kinase (FAK).
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IHC-P, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human FAK aa 700-800.

  • Positive control
    • WB and ICC: HeLa IHC: human hepatocellular carcinoma
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab40794 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/250 - 1/500.
IHC-P 1/250.
WB 1/1000. Detects a band of approximately 125 kDa (predicted molecular weight: 119 kDa).Can be blocked with Human FAK peptide (ab211922).
IP 1/50.

Target

  • Function
    Non-receptor protein-tyrosine kinase implicated in signaling pathways involved in cell motility, proliferation and apoptosis. Activated by tyrosine-phosphorylation in response to either integrin clustering induced by cell adhesion or antibody cross-linking, or via G-protein coupled receptor (GPCR) occupancy by ligands such as bombesin or lysophosphatidic acid, or via LDL receptor occupancy. Microtubule-induced dephosphorylation at Tyr-397 is crucial for the induction of focal adhesion disassembly. Plays a potential role in oncogenic transformations resulting in increased kinase activity.
  • Tissue specificity
    Expressed in all organs tested, in lymphoid cell lines, but most abundantly in brain.
  • Sequence similarities
    Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily.
    Contains 1 FERM domain.
    Contains 1 protein kinase domain.
  • Domain
    The first Pro-rich domain interacts with the SH3 domain of CRK-associated substrate (BCAR1) and CASL.
    The carboxy-terminal region is the site of focal adhesion targeting (FAT) sequence which mediates the localization of FAK1 to focal adhesions.
  • Post-translational
    modifications
    Phosphorylated on 6 tyrosine residues upon activation. Microtubule-induced dephosphorylation at Tyr-397 could be catalyzed by PTPN11 and regulated by ZFYVE21. Dephosphorylated by PTPN11 upon EPHA2 activation by its ligand EFNA1.
  • Cellular localization
    Cell junction > focal adhesion. Cell membrane. Constituent of focal adhesions.
  • Information by UniProt
  • Database links
  • Alternative names
    • FADK 1 antibody
    • FADK antibody
    • FAK related non kinase polypeptide antibody
    • FAK1 antibody
    • FAK1_HUMAN antibody
    • Focal adhesion kinase 1 antibody
    • Focal adhesion Kinase antibody
    • Focal adhesion kinase isoform FAK Del33 antibody
    • Focal adhesion kinase related nonkinase antibody
    • FRNK antibody
    • p125FAK antibody
    • pp125FAK antibody
    • PPP1R71 antibody
    • Protein phosphatase 1 regulatory subunit 71 antibody
    • Protein tyrosine kinase 2 antibody
    • Protein-tyrosine kinase 2 antibody
    • Ptk2 antibody
    • PTK2 protein tyrosine kinase 2 antibody
    see all

Images

  • ab40794 staining FAK in PANC-1 cells treated with CCK Octapeptide sulfated (ab120209), by ICC/IF. Increase of FAK expression correlates with increased concentration of CCK Octapeptide sulfated, as described in literature.
    The cells were incubated at 37°C for 10 minutes in media containing different concentrations of ab120209 (CCK Octapeptide sulfated) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab40794 (1/200) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • Anti-FAK antibody [EP695Y] (ab40794) at 1/1000 dilution + Hela cell lysate

    Predicted band size : 119 kDa
    Observed band size : 119 kDa
  • Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using ab40794.
  • Immunofluorescent staining of HeLa cells using ab40794.
  • The image shows FAK antibody (ab40794) in human spleen tissue. Clear cytoplasmic positivity in a subset of germinal centre cells. The there is intense positivity of the serum in the blood vessels. Endogenous peroxidases was blocked using 2% H2O2, for 15 minutes.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab40794 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40794, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.

  • All lanes : Anti-FAK antibody [EP695Y] (ab40794)

    Lane 1 : Milk PBS Tween
    Lane 2 : Milk PBS Tween

    Blocking peptides at 5 % per lane.

    Secondary
    HRP conjugated goat anti-rabbit poly clonal at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 119 kDa
    Observed band size : 130 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 minutes

    This image is courtesy of an Abreview submitted by Magali Boissiere (6970246)

    Western blot analysis of RAW264.7 cells lysate (40μg/lane) labelling FAK with ab40794 at 1/5000 in 5% Milk PBS Tween for 16 hours at 4ºC. A HRP conjugated goat anti-rabbit poly clonal (1/5000) was used as the secondary antibody.

     

    See Abreview

References

This product has been referenced in:
  • McKay TB  et al. Acute hypoxia influences collagen and matrix metalloproteinase expression by human keratoconus cells in vitro. PLoS One 12:e0176017 (2017). WB ; Human . Read more (PubMed: 28426715) »
  • Zhou C  et al. The Role of Titanium Surface Microtopography on Adhesion, Proliferation, Transformation, and Matrix Deposition of Corneal Cells. Invest Ophthalmol Vis Sci 57:1927-38 (2016). Read more (PubMed: 27092719) »

See all 26 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (RAW264.7 cells)
Gel Running Conditions
Reduced Denaturing (Gradient 4 -20%)
Loading amount
40 µg
Specification
RAW264.7 cells
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Magali Boissiere

Verified customer

Submitted Feb 23 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
(agent) for 10 minute(s) · Concentration: 3% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA, pH8.6, 100C, 20min
Sample
Human Tissue sections (placenta)
Specification
placenta
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 12 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Cow Cell (MDBK)
Specification
MDBK
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% Triton in PBS (20 min)
Blocking step
BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Nov 05 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Cow Cell lysate - whole cell (MDBK)
Loading amount
5 µg
Specification
MDBK
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Nov 02 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (hippocampus)
Loading amount
25 µg
Specification
hippocampus
Gel Running Conditions
Reduced Denaturing (6%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted May 31 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil: reactive)
Specification
Tonsil: reactive
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Jul 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Cor 1 cells - unstimulated)
Specification
Cor 1 cells - unstimulated
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% TWEEN 20
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Jul 19 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Jul 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HUVEC)
Specification
HUVEC
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% triton X-100
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 20 2010

Thank you for your enquiry. Unfortunately, the exact amino acid sequence information is proprietary and can not be released. However, I can provide you with a 50 amino acid sequence range from where the actual peptide sequence is located within, as ...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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