Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [CLIP/3127R] to CD74 - BSA and Azide free
  • Suitable for: WB, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free
    See all CD74 primary antibodies

  • Description

    Rabbit monoclonal [CLIP/3127R] to CD74 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt, IHC-Pmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Tissue, cells or virus. SU-DHL-4 lymphoma cells.

  • Positive control

    • IHC-P: Human tonsil tissue. WB: Raji and HEK-293T cell lysate. Flow Cyt: Raji cells.

  • General notes

    ab270296 is a carrier free version of ab270265.

     

    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

  • 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: 7.2
    Constituent: PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A/G purified

  • Purification notes

    Purified from bioreactor concentrate.

  • Clonality

    Monoclonal

  • Clone number

    CLIP/3127R

  • Isotype

    IgG

  • Research areas

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)

    Formalin-fixed, paraffin-embedded human tonsil tissue stained for CD74 using ab270265 at 2 μg/ml in immunohistochemical analysis. 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab270265).

  • Flow Cytometry - Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)
    Flow Cytometry - Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)

    This data was developed using the same antibody clone in a different buffer formulation (ab270265)

    Flow cytometry overlay histogram showing wild-type Raji (green line) and CD74 knockout Raji cells (ab273378) stained with ab270265 (red line). The cells were incubated in 1x PBS containing 10μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab270265) (1x106 in 100μl at 1 μg/ml) for 30 min at 4°C.

    The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150081) was used at 1/2000 for 30 min at 4°C.

    Isotype control antibody was Rabbit IgG (monoclonal) (ab172730) used at the same concentration and conditions as the primary antibody (wild-type Raji cells - black line; CD74 knockout Raji cells ab273378 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

  • Western blot - Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)
    Western blot - Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)
    All lanes : Anti-CD74 antibody [CLIP/3127R] (ab270265) at 1 µg/ml

    Lane 1 : Wild-type Raji cell lysate
    Lane 2 : CD74 knockout Raji cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : HepG2 cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 33-41 kDa
    Observed band size: 35 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab270265).

    Lanes 1 - 4: Merged signal (red and green). Green - ab270265 observed at 35 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab270265 was shown to react with CD74 in western blot. The band observed in CD74 knockout cell line ab273378 (knockout lysate ab275529) below 35 kDa is likely to represent a truncated form. This has not been investigated further. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab270265 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)
    Western blot - Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)
    Anti-CD74 antibody [CLIP/3127R] (ab270265) at 2 µg/ml + HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate

    Predicted band size: 33-41 kDa



    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab270265).

  • Flow Cytometry - Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)
    Flow Cytometry - Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)

    Flow cytometric analysis of Raji (Human Burkitt's lymphoma cell line) cells labeling CD74 using ab270265 at 2µg/106 cells followed by goat anti-mouse IgG-CF488 (Blue); isotype control (Red).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab270265).

  • Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)
    Anti-CD74 antibody [CLIP/3127R] - BSA and Azide free (ab270296)

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