Overview

  • Product name
    Anti-Src antibody [Clone 327]
    See all Src primary antibodies
  • Description
    Mouse monoclonal [Clone 327] to Src
  • Specificity
    Reacts with both v-Src and c-Src proteins.
  • Tested applications
    Suitable for: Flow Cyt, WB, IPmore details
    Unsuitable for: ICC/IF,IHC-Fr or IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Full length native protein (purified) corresponding to Src. Antibody generated by immunizing BALB/c mice with full length native protein (purified) and then fusing with P3X63 Ag8.653 myeloma cells.

  • Epitope
    Epitope is within the SH3 domain of either v-Src or c-Src. We have no evidence for ab16885 cross-reacting with other src family kinases (e.g.fyn).
  • Positive control
    • WB: MCF7, HeLa, A431 and NIH/3T3 cell lysates. Flow Cyt: SH-SY5Y cells.
  • General notes

    May appear as a doublet due to phosphorylation. May be used to precipitate active Src which can then be used in a kinase reaction.

Properties

Applications

Our Abpromise guarantee covers the use of ab16885 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 Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB Use a concentration of 2.5 µg/ml. Predicted molecular weight: 59.7 kDa. ab16885 may appear as a doublet on WB due to phosphorylation. This antibody can be used to precipitate active src kinase.
IP Use at 5 µg/mg of lysate. See Abreview.
  • Application notes
    Is unsuitable for ICC/IF,IHC-Fr or IHC-P.
  • Target

    • Function
      Non-receptor protein tyrosine kinase that plays pivotal roles in numerous cellular processes such as proliferation, migration, and transformation. In concert with PTK2B, plays an important role in osteoclastic bone resorption. Both the formation of a SRC-PTK2B complex, and SRC kinase activity are necessary for this function. Once it is recruited to the activated integrins, by PTK2B, it phosphorylates CBL which in turn induces the activation and recruitment of phosphatidylinositol 3-kinase to the cell membrane in a signaling pathway that is critical for osteoclast function. Promotes energy production in osteoclasts by activating mitochondrial cytochrome C oxidase. Phosphorylates RUNX3 and COX2 on tyrosine residues, TNK2 on 'Tyr-284' and CBL on 'Tyr-731'. Enhances DDX58/RIG-I-elicited antiviral signaling.
    • Sequence similarities
      Belongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.
      Contains 1 protein kinase domain.
      Contains 1 SH2 domain.
      Contains 1 SH3 domain.
    • Post-translational
      modifications
      Dephosphorylated at Tyr-530 by PTPRJ (By similarity). Phosphorylated on Tyr-530 by c-Src kinase (CSK). The phosphorylated form is termed pp60c-src. Dephosphorylated by PTPRJ at Tyr-419. Normally maintained in an inactive conformation with the SH2 domain engaged with Tyr-530, the SH3 domain engaged with the SH2-kinase linker, and Tyr-419 dephosphorylated. Dephosphorylation of Tyr-530 as a result of protein tyrosine phosphatase (PTP) action disrupts the intramolecular interaction between the SH2 domain and Tyr-530, Tyr-419 can then become autophosphorylated, resulting in SRC activation. Phosphorylation of Tyr-530 by CSK allows this interaction to reform, resulting in SRC inactivation.
      S-nitrosylation is important for activation of its kinase activity.
    • Cellular localization
      Cell membrane. Mitochondrion inner membrane.
    • Information by UniProt
    • Database links
    • Form
      This protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), FYN (P06241), YES1 (P07947), and LYN (P07948). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request.
    • Alternative names
      • ASV antibody
      • Avian sarcoma virus antibody
      • c SRC antibody
      • CDNA FLJ14219 fis clone NT2RP3003800 highly similar to Rattus norvegicus tyrosine protein kinase pp60 c src mRNA antibody
      • cSrc antibody
      • EC 2.7.10.2 antibody
      • Neuronal CSRC tyrosine specific protein kinase antibody
      • Neuronal SRC antibody
      • Oncogene SRC antibody
      • OTTHUMP00000174476 antibody
      • OTTHUMP00000174477 antibody
      • p60 Src antibody
      • p60-Src antibody
      • p60Src antibody
      • pp60c src antibody
      • pp60c-src antibody
      • pp60csrc antibody
      • Proto oncogene tyrosine protein kinase Src antibody
      • Proto-oncogene c-Src antibody
      • Proto-oncogene tyrosine-protein kinase Src antibody
      • Protooncogene SRC antibody
      • Protooncogene SRC Rous sarcoma antibody
      • Src antibody
      • SRC Oncogene antibody
      • SRC proto oncogene non receptor tyrosine kinase antibody
      • SRC_HUMAN antibody
      • SRC1 antibody
      • Tyrosine kinase pp60c src antibody
      • Tyrosine protein kinase SRC 1 antibody
      • Tyrosine protein kinase SRC1 antibody
      • v src avian sarcoma (Schmidt Ruppin A2) viral oncogene homolog antibody
      • V src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog (avian) antibody
      • v src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog avian antibody
      see all

    Anti-Src antibody [Clone 327] images



    • Predicted band size : 59.7 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Src knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: A431 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab16885 observed at 60 kDa. Red - loading control, ab8227, observed at 42 kDa.

      ab16885 was shown to specifically react with Src when Src knockout samples were used. Wild-type and Src knockout samples were subjected to SDS-PAGE. ab16885 and ab8227 (loading control to beta Actin) were diluted 2.5µg/ml and 1/1000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    • Overlay histogram showing SH-SY5Y cells stained with ab16885 (red line). The cells were fixed with 80% 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 (ab16885, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.



    • Predicted band size : 59.7 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Src knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: A431 cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab16885 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.

      This western blot image is a comparison between ab16885 and a competitor's top cited rabbit polyclonal antibody.

    • All lanes : Anti-Src antibody [Clone 327] (ab16885) at 2.5 µg/ml

      Lane 1 : Whole cell lysate prepared from human MCF-7 cells (Untreated)
      Lane 2 : Whole cell lysate prepared from human MCF-7 cells (Treated with 10uM PP2)

      Secondary
      HRP conjugated Donkey polyclonal to mouse Ig at 1/5000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 59.7 kDa


      Exposure time : 3 minutes

      This image is courtesy of an anonymous Abreview

      See Abreview

    • Anti-Src antibody [Clone 327] (ab16885) at 1/100 dilution + Lysate prepared from mouse NIH 3T3 cell line at 10 µg

      Secondary
      HRP-conjugated sheep monoclonal to mouse IgG at 1/5000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 59.7 kDa
      Observed band size : 60 kDa (why is the actual band size different from the predicted?)


      Exposure time : 12 minutes

      This image is a courtesy of Anonymous Abreview

      See Abreview

    References for Anti-Src antibody [Clone 327] (ab16885)

    This product has been referenced in:
    • Pérez Y  et al. Lipid binding by the Unique and SH3 domains of c-Src suggests a new regulatory mechanism. Sci Rep 3:1295 (2013). WB . Read more (PubMed: 23416516) »
    • Ablooglu AJ  et al. Antithrombotic effects of targeting alphaIIbbeta3 signaling in platelets. Blood 113:3585-92 (2009). WB ; Mouse . Read more (PubMed: 19005179) »

    See all 2 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (NIH 3T3 murine cell line)
    Loading amount
    10 µg
    Specification
    NIH 3T3 murine cell line
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted Mar 23 2010

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (HT1080 cells)
    Loading amount
    10 µg
    Specification
    HT1080 cells
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 22 2009

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (MCF7)
    Total protein in input
    1e+006 cells
    Specification
    MCF7
    Immuno-precipitation step
    Other - Dynabeads
    Username

    Abcam user community

    Verified customer

    Submitted Dec 17 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (MCF-7)
    Loading amount
    30 µg
    Specification
    MCF-7
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Dec 16 2008

    Thank you for your patience. Regarding ab16885 (Mouse monoclonal [Clone 327] to Src), this antibody has not been tested on mouse platelets or src-null cells. Also, we have not tested this product for crossreactivity with other src family members e....

    Read More

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

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