Key features and details
- Mouse monoclonal [PIN.1] to CD74
- Suitable for: IHC-P, ICC/IF, WB, IP, Flow Cyt
- Reacts with: Human
- Isotype: IgG1
Product nameAnti-CD74 antibody [PIN.1]
See all CD74 primary antibodies
DescriptionMouse monoclonal [PIN.1] to CD74
SpecificityThis antibody detects an ~33-35 kDa protein doublet, corresponding to the apparent molecular mass of the p33 and p35 forms of human CD74.
Tested applicationsSuitable for: IHC-P, ICC/IF, WB, IP, Flow Cytmore details
Species reactivityReacts with: Human
EpitopeThe epitope is in a region of the cytoplasmic tail of human CD74 which is common to all isoforms.
- Human B lymphoblastoid cell lysate. IF/ICC: HepG2 cell line.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.09% Sodium azide
Constituents: PBS, 50% Glycerol
Concentration information loading...
PurityProtein G purified
Purification notesThis antibody was purified by immunoaffinity chromatography.
Our Abpromise guarantee covers the use of ab22603 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 10 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 33 kDa.|
|IP||Use a concentration of 12 µg/ml.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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
Ab22603 staining CD74 in human liver. Staining is localized to the membrane and cytoplasm.
Left panel: with primary antibody at 4ug/ml. Right panel: isotype control.
Sections were stained using an automated system (Dako Autostainer plus), at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer, EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be req
ICC/IF image of ab22603 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab22603, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM
ICC/IF image of ab22603 staining CD74 in HaCaT cells. Cells were fixed with cold 100% methanol for 10 minutes at -20°C. Samples were incubated with primary antibody at 1:100 dilution for 1 hour at room temperature. A FITC Goat anti-mouse (green) was used as a secondary antibody at 1:50 dilution for 1 hour at room temperature.
Overlay histogram showing Raji cells stained with ab22603 (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. The cells were then incubated with the antibody (ab22603, 1µg/1x106 cells) 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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Raji cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.
ab22603 has been referenced in 7 publications.
- Kong XF et al. Disruption of an antimycobacterial circuit between dendritic and helper T cells in human SPPL2a deficiency. Nat Immunol 19:973-985 (2018). PubMed: 30127434
- Tato M et al. Cathepsin S inhibition combines control of systemic and peripheral pathomechanisms of autoimmune tissue injury. Sci Rep 7:2775 (2017). PubMed: 28584258
- Theron M et al. Pharmacodynamic Monitoring of RO5459072, a Small Molecule Inhibitor of Cathepsin S. Front Immunol 8:806 (2017). WB ; Human . PubMed: 28769925
- van de Weijer ML et al. The E3 Ubiquitin Ligase TMEM129 Is a Tri-Spanning Transmembrane Protein. Viruses 8:N/A (2016). Human . PubMed: 27854284
- Fernandez-Cuesta L et al. CD74-NRG1 Fusions in Lung Adenocarcinoma. Cancer Discov 4:415-22 (2014). PubMed: 24469108
- Landsverk OJ et al. Invariant chain increases the half-life of MHC II by delaying endosomal maturation. Immunol Cell Biol 89:619-29 (2011). PubMed: 21116285
- Leth-Larsen R et al. Metastasis-related plasma membrane proteins of human breast cancer cells identified by comparative quantitative mass spectrometry. Mol Cell Proteomics 8:1436-49 (2009). PubMed: 19321434