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
Product nameAnti-CD74 antibody [EPR4064] - BSA and Azide free
See all CD74 primary antibodies
DescriptionRabbit monoclonal [EPR4064] to CD74 - BSA and Azide free
Tested applicationsSuitable for: ICC, Flow Cyt (Intra), IHC-P, Flow Cyt, WBmore details
Unsuitable for: IP
Species reactivityReacts with: Human
Synthetic peptide within Human CD74 aa 1-100. The exact sequence is proprietary.
Database link: P04233
- Raji and Ramos cell lysate; Human tonsil tissue
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
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.
Storage instructionsShipped at 4°C. Store at +4°C. Do Not Freeze.
Storage bufferpH: 7.2
Concentration information loading...
PurityProtein A purified
KO cell lines
KO cell lysates
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.
|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.|
FunctionPlays 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 similaritiesContains 1 thyroglobulin type-1 domain.
Cellular localizationCell 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
- CD 74 antibody
- CD74 antibody
- CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated) antibody
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).
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.
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.
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.
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.
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).
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.
ab247655 has not yet been referenced specifically in any publications.