Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)

Submit a review Submit a question
Western blot - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Flow Cytometry (Intracellular) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Immunocytochemistry - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Flow Cytometry - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Western blot - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
  • Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4064] to CD74 - BSA and Azide free
  • Suitable for: ICC, Flow Cyt (Intra), IHC-P, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-CD74 antibody [EPR4064] - BSA and Azide free
    See all CD74 primary antibodies

  • Description

    Rabbit monoclonal [EPR4064] to CD74 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, Flow Cyt (Intra), IHC-P, Flow Cyt, WBmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human CD74 aa 1-100. The exact sequence is proprietary.
    Database link: P04233

  • Positive control

    • Raji and Ramos cell lysate; Human tonsil tissue

  • General notes

    ab247655 is the carrier-free version of ab108393 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab247655 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Storage buffer

    pH: 7.2
    Constituent: PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR4064

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab247655 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
ICC Use at an assay dependent concentration.
Flow Cyt (Intra) Use a concentration of 0.2 µg/ml.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Perform antigen retrieval before commencing with IHC staining protocol.

 
Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 34 kDa.
  • Application notes
    Is unsuitable for IP.

    • Target

    • Function

      Plays a critical role in MHC class II antigen processing by stabilizing peptide-free class II alpha/beta heterodimers in a complex soon after their synthesis and directing transport of the complex from the endoplasmic reticulum to the endosomal/lysosomal system where the antigen processing and binding of antigenic peptides to MHC class II takes place. Serves as cell surface receptor for the cytokine MIF.

    • Sequence similarities

      Contains 1 thyroglobulin type-1 domain.

    • Cellular localization

      Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus > trans-Golgi network. Endosome. Lysosome. Transits through a number of intracellular compartments in the endocytic pathway. It can either undergo proteolysis or reach the cell membrane.
    • Information by UniProt

    • Database links

      • Alternative names

        • CD 74 antibody
        • CD74 antibody
        • CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated) antibody
        • CD74 antigen antibody
        • CD74 molecule antibody
        • CD74 molecule, major histocompatibility complex, class II invariant chain antibody
        • CLIP antibody
        • DHLAG antibody
        • Gamma chain of class II antigens antibody
        • HG2A_HUMAN antibody
        • HLA class II histocompatibility antigen gamma chain antibody
        • HLA DR antigens associated invariant chain antibody
        • HLA DR gamma antibody
        • HLA-DR antigens-associated invariant chain antibody
        • HLA-DR-gamma antibody
        • HLADG antibody
        • HLADR antigens associated invariant chain antibody
        • Ia antigen associated invariant chain antibody
        • Ia antigen-associated invariant chain antibody
        • Ia associated invariant chain antibody
        • Ia gamma antibody
        • Ii antibody
        • Invariant polypeptide of major histocompatibility complex class II antigen associated antibody
        • la-gamma antibody
        • Major histocompatibility complex class II invariant chain antibody
        • MHC HLA DR gamma chain antibody
        • MHC HLA-DR gamma chain antibody
        • p33 antibody
        • p35 antibody
        • Protein 41 antibody
        see all

      Images

      • Western blot - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Western blot - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        All lanes : Anti-CD74 antibody [EPR4064] (ab108393) at 1/1000 dilution

        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: 34 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 (ab108393).

        Lanes 1 - 4: Merged signal (red and green). Green - ab108393 observed at 35 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

        ab108393 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 fluorescent western blot (TBS-based) blocking solution before incubation with ab108393 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 h at room temperature before imaging.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)

        This data was developed using ab108393, the same antibody clone in a different buffer formulation.

        Immunohistochemical analysis of CD74 in paraffin embedded Human tonsil tissue, using ab108393 at a 1/100 dilution. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
      • Flow Cytometry (Intracellular) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Flow Cytometry (Intracellular) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)

        This data was developed using the same antibody clone in a different buffer formulation (ab108393)

        Flow cytometry overlay histogram showing wild-type Raji (green line) and CD74 knockout Raji cells (ab273378) stained with ab108393 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then 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 (ab108393) (1x106 in 100μl at 0.2 μg/ml) for 30 min at 22°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 22°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.

      • Immunocytochemistry - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Immunocytochemistry - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)

        This data was developed using ab108393, the same antibody clone in a different buffer formulation.

        Confocal image of ab108393 staining CD74 in the RAMOS (human Burkitt's lymphoma) cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100 in PBS. Samples were incubated with primary antibody (1/500) and an Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used ad a nuclear counterstain at 1/200.
      • Flow Cytometry - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Flow Cytometry - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)

        This data was developed using ab108393, the same antibody clone in a different buffer formulation.

        ab108393 staining CD74 in RAMOS (human Burkitt's lymphoma) cell line. The sample was fixed with 4% paraformaldehyde and incubated with the primary antibody (1/150) for 30 minutes at 4°C. An Alexa Fluor® 488 -conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Control: Cell without incubation with primary antibody and secondary antibody (Blue).
      • Western blot - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Western blot - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Anti-CD74 antibody [EPR4064] (ab108393) at 1/1000 dilution + Ramos cell lysate at 10 µg

        Predicted band size: 34 kDa



        This data was developed using ab108393, the same antibody clone in a different buffer formulation.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)

        This data was developed using ab108393, the same antibody clone in a different buffer formulation.

        ab108393 showing negative staining in Normal brain tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        This data was developed using ab108393, the same antibody clone in a different buffer formulation.ab108393 showing positive staining in B cell lymphoma tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        This data was developed using ab108393, the same antibody clone in a different buffer formulation.ab108393 showing negative staining in Normal placenta tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        This data was developed using ab108393, the same antibody clone in a different buffer formulation.ab108393 showing positive staining in Normal spleen tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        This data was developed using ab108393, the same antibody clone in a different buffer formulation.ab108393 showing negative staining in Normal heart tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        This data was developed using ab108393, the same antibody clone in a different buffer formulation.ab108393 showing negative staining in Normal liver tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
      • Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)
        Anti-CD74 antibody [EPR4064] - BSA and Azide free (ab247655)

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References (0)

      Publishing research using ab247655? Please let us know so that we can cite the reference in this datasheet.

      ab247655 has not yet been referenced specifically in any publications.

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      There are currently no Customer reviews or Questions for ab247655.
      Please use the links above to contact us or submit feedback about this product.

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com