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Anti-Src antibody [Clone 327] (ab16885)

Key features and details

  • Mouse monoclonal [Clone 327] to Src
  • Suitable for: Flow Cyt, WB
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-Src antibody [Clone 327]
    See all Src primary antibodies

  • Description

    Mouse monoclonal [Clone 327] to Src

  • Host species

    Mouse

  • Specificity

    Reacts with both v-Src and c-Src proteins.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    WB
    Human
    See all applications and species data

  • 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. Wild-type HAP1 cell lysate. 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.

    This product was changed from ascites to tissue culture supernatant on 17 May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Images

  • Western blot - Anti-Src antibody [Clone 327] (ab16885)
    Western blot - Anti-Src antibody [Clone 327] (ab16885)

    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.

    This image was generated using the ascites version of the product.

  • Flow Cytometry - Anti-Src antibody [Clone 327] (ab16885)
    Flow Cytometry - Anti-Src antibody [Clone 327] (ab16885)

    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.

    This image was generated using the ascites version of the product.

  • Western blot - Anti-Src antibody [Clone 327] (ab16885)
    Western blot - Anti-Src antibody [Clone 327] (ab16885)

    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.

    This image was generated using the ascites version of the product.

  • Western blot - Anti-Src antibody [Clone 327] (ab16885)
    Western blot - Anti-Src antibody [Clone 327] (ab16885) This image is courtesy of an anonymous Abreview
    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
    All lanes : 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 was generated using the ascites version of the product.

    See Abreview

  • Western blot - Anti-Src antibody [Clone 327] (ab16885)
    Western blot - Anti-Src antibody [Clone 327] (ab16885) This image is a courtesy of Anonymous 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 was generated using the ascites version of the product.

    See Abreview

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