Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR8569] to CD27 - Low endotoxin, Azide free
  • Suitable for: WB, ICC/IF, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free
    See all CD27 primary antibodies

  • Description

    Rabbit monoclonal [EPR8569] to CD27 - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, IHC-P, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Raji, Ramos and lymph node cell lysates; Human stomach and tonsil tissues.

  • General notes

    ab192336 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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

  • Flow Cytometry - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    Flow Cytometry - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)

    Flow Cytometry analysis of Ramos cells labelling CD27 with purified ab131254 at 1/300 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).

  • Western blot - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    Western blot - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    All lanes : Anti-CD27 antibody [EPR8569] (ab131254) at 1/1000 dilution

    Lane 1 : Wild-type Raji cell lysate
    Lane 2 : CD27 knockout Raji cell lysate
    Lane 3 : Ramos cell lysate
    Lane 4 : Human brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 29 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 (ab131254).

    Lanes 1 - 4: Merged signal (red and green). Green - ab131254 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

    ab131254 was shown to react with CD27 in wild-type Raji cells in western blot with loss of signal observed in CD27 knockout sample. Wild-type and CD27 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab131254 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution 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 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD27 with purified ab131254 at 1/1500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue showing negative staining with purified ab131254 at 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).

  • Immunocytochemistry/ Immunofluorescence - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    Immunocytochemistry/ Immunofluorescence - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)

    Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling CD27 with purified ab131254 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

    Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)

    Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections) analysis of human stomach tissue labelling CD27 with ab131254 at a dilution of 1/100.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)

    Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections) analysis of human tonsil tissue labelling CD27 with ab131254 at a dilution of 1/100.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • OI-RD Scanning - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    OI-RD Scanning - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).

  • Western blot - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    Western blot - Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336) + Human spleen lysate at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

    Predicted band size: 29 kDa
    Observed band size: 55 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM/TBST

  • Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
    Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)

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