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    products/primary-antibodies/fak-antibody-ep1831y-ab76496.pdf

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Cardiovascular Angiogenesis Adhesion / ECM Integrins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-FAK antibody [EP1831Y] (ab76496)

  • Datasheet
  • SDS
Submit a review Q&A (3)References (4)

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Western blot - Anti-FAK antibody [EP1831Y] (ab76496)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAK antibody [EP1831Y] (ab76496)
  • Western blot - Anti-FAK antibody [EP1831Y] (ab76496)
  • Anti-FAK antibody [EP1831Y] (ab76496)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1831Y] to FAK
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

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Human PTK2 (FAK) knockout HEK-293T cell line (ab255421)

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Overview

  • Product name

    Anti-FAK antibody [EP1831Y]
    See all FAK primary antibodies
  • Description

    Rabbit monoclonal [EP1831Y] to FAK
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human FAK aa 300-400. The exact sequence is proprietary.

  • Positive control

    • WB: HeLa, A431 and HEK-293T cell lysate. IHC-P: human tonsil tissue.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1831Y
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Angiogenesis
    • Adhesion / ECM
    • Integrins
    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • FAK / PYK
    • Signal Transduction
    • Cytoskeleton / ECM
    • Extracellular Matrix
    • Structures
    • Focal Adhesions
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Tyrosine kinases
    • FAK family

Associated products

  • Alternative Versions

    • Anti-FAK antibody [EP1831Y] - BSA and Azide free (ab247481)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human PTK2 (FAK) knockout HEK-293T cell line (ab255421)
  • KO cell lysates

    • Human PTK2 (FAK) knockout HEK-293T cell lysate (ab263766)
  • Recombinant Protein

    • Recombinant human FAK protein (ab105909)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab76496 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
WB
1/500 - 1/2000. Predicted molecular weight: 119 kDa.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Notes
WB
1/500 - 1/2000. Predicted molecular weight: 119 kDa.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Application notes
Is unsuitable for IP.

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.
  • Target information above from: UniProt accession Q05397 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5747 Human
    • Entrez Gene: 14083 Mouse
    • Entrez Gene: 25614 Rat
    • Omim: 600758 Human
    • SwissProt: Q05397 Human
    • SwissProt: P34152 Mouse
    • SwissProt: O35346 Rat
    • Unigene: 395482 Human
    • Unigene: 254494 Mouse
    • Unigene: 2809 Rat
    see all
  • 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

  • Western blot - Anti-FAK antibody [EP1831Y] (ab76496)
    Western blot - Anti-FAK antibody [EP1831Y] (ab76496)
    All lanes : Anti-FAK antibody [EP1831Y] (ab76496) at 1/500 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : Wild-type HEK-293T cell lysate
    Lane 4 : PTK2 knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 119 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab76496 observed at 119 kDa. Red - loading control, ab8245 observed at 37 kDa.

     ab76496 was shown to react with FAK in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255421 (knockout cell lysate ab263766) was used. Wild-type and FAK knockout samples were subjected to SDS-PAGE. ab76496 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAK antibody [EP1831Y] (ab76496)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAK antibody [EP1831Y] (ab76496)

    Immunohistochemical staining of paraffin-embedded human tonsils using 1/100 ab76496

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-FAK antibody [EP1831Y] (ab76496)
    Western blot - Anti-FAK antibody [EP1831Y] (ab76496)
    Anti-FAK antibody [EP1831Y] (ab76496) at 1/500 dilution + HeLa cell lysate at 10 µg

    Secondary
    HRP labeled goat anti-rabbit at 1/1000 dilution

    Predicted band size: 119 kDa

  • Anti-FAK antibody [EP1831Y] (ab76496)
    Anti-FAK antibody [EP1831Y] (ab76496)

Protocols

  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (4)

Publishing research using ab76496? Please let us know so that we can cite the reference in this datasheet.

ab76496 has been referenced in 4 publications.

  • Husari A  et al. On the relationship of YAP and FAK in hMSCs and osteosarcoma cells: Discrimination of FAK modulation by nuclear YAP depletion or YAP silencing. Cell Signal 63:109382 (2019). PubMed: 31376525
  • Cui Y & Yang S Overexpression of Annexin A1 protects against benzo[a]pyrene-induced bronchial epithelium injury. Mol Med Rep 18:349-357 (2018). PubMed: 29749523
  • Lu D  et al. MicroRNA-146a promote cell migration and invasion in human colorectal cancer via carboxypeptidase M/src-FAK pathway. Oncotarget 8:22674-22684 (2017). WB ; Human . PubMed: 28186967
  • Goodwin JM  et al. An AMPK-independent signaling pathway downstream of the LKB1 tumor suppressor controls Snail1 and metastatic potential. Mol Cell 55:436-50 (2014). PubMed: 25042806

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-3 of 3 Abreviews or Q&A

Question

Human melanoma tumor cells line and HT29 colon cancer cell line used
ab4803
antibody shows multiple bands
20ug to 200ug protein loaded
used at 1/200, 1/500 and 1/1000 dilution
Milk and BSA used for blocking overnight
Lysates heated 55C for 10 minutes with loading buffer

ab76496
fainnt band
20-200ug protein loaded
1/200, 1/500 and 1/1000 dilution
Surprisingly 1/500 gave high intensity band than 1/200
Used BSA milk
Lysates heated 55C for 10 minutes with loading buffer

Read More

Abcam community

Verified customer

Asked on Jul 31 2012

Answer

According to our records, ab76496 and ab4803was proving difficult to use inWB and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

We wish you the best of luck with your research and look forward to a reply.

Read More

Abcam Scientific Support

Answered on Jul 31 2012

Question

Product code: 76496
Lot number: GR48341-2

Inquiry: I Had problem with both antibodies and I send you a summary of most of conditions tried. other conditions such as overnight blocking but the result was the same no band present.

Read More

Abcam community

Verified customer

Asked on May 29 2012

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that ab76496 is tested and covered by our 6 month guarantee forwestern blot and for Mouse, Rat andHuman samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you could also provide an image which would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionaire.


Order Details
Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

Read More

Abcam Scientific Support

Answered on May 29 2012

Question

Human melanoma tumor cells line and HT29 colon cancer cell line used
ab4803
antibody shows multiple bands
20ug to 200ug protein loaded
used at 1/200, 1/500 and 1/1000 dilution
Milk and BSA used for blocking overnight
Lysates heated 55C for 10 minutes with loading buffer

ab76496
fainnt band
20-200ug protein loaded
1/200, 1/500 and 1/1000 dilution
Surprisingly 1/500 gave high intensity band than 1/200
Used BSA milk
Lysates heated 55C for 10 minutes with loading buffer

Read More

Abcam community

Verified customer

Asked on May 18 2012

Answer

Thank you for contacting us.

Please repeat the experiment as we discussed on phone,If the results does not improve please do not hesitate to contact me for further assistance.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Abcam Scientific Support

Answered on May 18 2012

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