Overview

  • Product name
    Anti-Cortactin (phospho Y421) antibody
    See all Cortactin primary antibodies
  • Description
    Rabbit polyclonal to Cortactin (phospho Y421)
  • Host species
    Rabbit
  • Specificity
    This antibody detects Cortactin only when phosphorylated at tyrosine 421.
  • Tested applications
    Suitable for: WB, ELISAmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthesized phosphopeptide derived from human Cortactin around the phosphorylation site of tyrosine 421 (P-I-YP-E-D).

  • Positive control
    • Hela cell extract.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride

    Without Mg+2 and Ca+2
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab47768 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 - 1/1000. Detects a band of approximately 85 kDa (predicted molecular weight: 62 kDa).
ELISA 1/20000.

Target

  • Function
    Contributes to the organization of the actin cytoskeleton and cell structure. Plays a role in the regulation of cell migration. Plays a role in the invasiveness of cancer cells, and the formation of metastases.
  • Sequence similarities
    Contains 7 cortactin repeats.
    Contains 1 SH3 domain.
  • Domain
    The SH3 motif may mediate binding to the cytoskeleton.
  • Post-translational
    modifications
    Tyrosine phosphorylation in transformed cells may contribute to cellular growth regulation and transformation.
  • Cellular localization
    Cytoplasm > cytoskeleton. Cell projection > lamellipodium. Cell projection > ruffle. Associated with membrane ruffles and lamellipodia.
  • Information by UniProt
  • Database links
  • Alternative names
    • Amplaxin antibody
    • CTTN antibody
    • EMS 1 antibody
    • EMS1 antibody
    • FLJ34459 antibody
    • Mammary tumor and squamous cell carcinoma associated antibody
    • Oncogene EMS1 antibody
    • p80/85 src substrate antibody
    • Src substrate cortactin antibody
    • SRC8_HUMAN antibody
    see all

Images

  • All lanes : Anti-Cortactin (phospho Y421) antibody (ab47768) at 1/500 dilution

    Lane 1 : Hela cell extract, treated with H2O2
    Lane 2 : Hela cell extract, untreated

    Predicted band size: 62 kDa



    Typically 5-30ug of total protein was loaded per lane of the gel. The cells were treated with 150 uM H2O2.

References

This product has been referenced in:
  • Meiler E  et al. Cortactin tyrosine phosphorylation promotes its deacetylation and inhibits cell spreading. PLoS One 7:e33662 (2012). WB . Read more (PubMed: 22479425) »
  • García-Castillo J  et al. HER2 carboxyl-terminal fragments regulate cell migration and cortactin phosphorylation. J Biol Chem 284:25302-13 (2009). WB ; Human . Read more (PubMed: 19589785) »
See all 2 Publications for this product

Customer reviews and Q&As

Answer

Thank you for contacting us. I am sorry to hear that this antibody is not providing satisfactory results.

Would you be able to provide the lot number and order number?

It may be possible that the ˜100 kDa band corresponds to the phosphorylated form of cortactin. Since the cortactin antibody you're using ab33333 is a monoclonal with one specific epitope, it may not have an epitope that encompasses every phosphorylation site (24 possible from Swiss Prot) present on cortactin to make it a true indicator of total cortactin (meaning phospho- and non-). Plus phosphoproteins run higher in gels, so it seems to fit where we may expect the band to arise.

Also, are you treating your cells to induce phosphorylation at this particular site (Y421)? Our positive control was HeLa cells treated with 150 uM H2O2. Is it known the squamous cell carcinomas endogenously express this phosphorylation? The best information I could find online was with a serum starved squamous cell carcinoma of the oropharynx. Were your cells serum starved? http://www.phosphosite.org/siteAction.do?id=2685

Lastly, if you are blocking in milk, I'd recommend switching to 5% BSA throughout the entire experiment since milk contains endogenous phosphatases.

I hope this information helps. I look forward to your reply so that we may help you further.

Read More

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

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