Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RIP antibody [7H10] (ab72139)

Key features and details

  • Mouse monoclonal [7H10] to RIP
  • Suitable for: Flow Cyt, WB, IHC-P
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG2b

Overview

  • Product name

    Anti-RIP antibody [7H10]
    See all RIP primary antibodies

  • Description

    Mouse monoclonal [7H10] to RIP

  • Host species

    Mouse

  • Tested Applications & Species

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

  • Immunogen

    Recombinant full length human RIP protein purified from E.coli (His/ABD-RIP)

  • Positive control

    • HeLa, K562 and SK-N-MC cell lysates

  • General notes

    This product was changed from ascites to tissue culture supernatant on 18th September 2017. Lot numbers higher than GR304252 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-RIP antibody [7H10] (ab72139)
    Western blot - Anti-RIP antibody [7H10] (ab72139)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: RIP knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: Raji cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab72139 observed at 78 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab72139 was shown to recognize RIP in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when RIP knockout samples were examined. Wild-type and RIP knockout samples were subjected to SDS-PAGE. ab72139 at a dilution of 1/500 and ab181602 (loading control to GAPDH) at a dilution of 1/10,000 were 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 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-RIP antibody [7H10] (ab72139)
    Western blot - Anti-RIP antibody [7H10] (ab72139)
    All lanes : Anti-RIP antibody [7H10] (ab72139) at 1/500 dilution

    Lane 1 : K562 cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : SK-N-MC cell lysate

    Predicted band size: 76 kDa
    Observed band size: 76 kDa
    Additional bands at: 57 kDa. We are unsure as to the identity of these extra bands.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RIP antibody [7H10] (ab72139)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RIP antibody [7H10] (ab72139)
    IHC image of ab72139 staining in Breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab72139, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • Flow Cytometry - Anti-RIP antibody [7H10] (ab72139)
    Flow Cytometry - Anti-RIP antibody [7H10] (ab72139)
    Overlay histogram showing HEK293 cells stained with ab72139 (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 (ab72139, 1/100 dilution) 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 IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

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