Mouse Monoclonal CD74 antibody. Suitable for IHC-P, Flow Cyt, WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 23 publications.
IgG1
Mouse
pH: 7.3 - 7.5
Preservative: 0.1% Sodium azide
Constituents: 1% BSA
Liquid
Monoclonal
IHC-P | Flow Cyt | WB | Flow Cyt (Intra) | |
---|---|---|---|---|
Human | Tested | Tested | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/25.00000 - 1/50.00000 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/10 | Notes Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 5 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
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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.Class-II-associated invariant chain peptideBinds to the peptide-binding site of MHC class II alpha/beta heterodimers forming an alpha-beta-CLIP complex, thereby preventing the loading of antigenic peptides to the MHC class II complex until its release by HLA-DM in the endosome.Isoform p41Stabilizes the conformation of mature CTSL by binding to its active site and serving as a chaperone to help maintain a pool of mature enzyme in endocytic compartments and extracellular space of antigen-presenting cells (APCs). Has antiviral activity by stymieing the endosomal entry of Ebola virus and coronaviruses, including SARS-CoV-2 (PubMed:32855215). Disrupts cathepsin-mediated Ebola virus glycoprotein processing, which prevents viral fusion and entry. This antiviral activity is specific to p41 isoform (PubMed:32855215).
DHLAG, CD74, DHLAG, HLA class II histocompatibility antigen gamma chain, HLA-DR antigens-associated invariant chain, Ia antigen-associated invariant chain, Ii
Mouse Monoclonal CD74 antibody. Suitable for IHC-P, Flow Cyt, WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 23 publications.
IgG1
Mouse
pH: 7.3 - 7.5
Preservative: 0.1% Sodium azide
Constituents: 1% BSA
Liquid
Monoclonal
LN2
Affinity purification Protein A/G
kappa
Blue Ice
1-2 weeks
+4°C
-20°C
Avoid freeze / thaw cycle
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This supplementary information is collated from multiple sources and compiled automatically.
CD74 also known as the invariant chain or li is a protein with a mass of about 33 kDa. It is widely expressed on the surface of immune cells such as B cells dendritic cells and macrophages. CD74 plays an important role in the immune system by acting as a chaperone for major histocompatibility complex class II (MHC II) molecules guiding their migration to endosomes where they encounter antigenic peptides. The CD74 protein also functions as a cell surface receptor for macrophage migration inhibitory factor (MIF) enhancing the immune response process.
CD74 facilitates multiple immune regulatory processes. It forms a complex with the MHC II molecules aiding in their proper folding and peptide loading in the endoplasmic reticulum. This complex then travels through the Golgi apparatus towards the lysosomal compartments where antigens are presented to initiate adaptive immune responses. In addition CD74 is involved in signal transduction pathways that regulate cell proliferation and survival influenced by interactions with MIF and other molecules.
The role of CD74 extends into both the antigen presentation and MIF signaling pathways. Within the antigen presentation pathway CD74's activity is tightly connected with MHC II and subsequently to CD4+ T cells facilitating the activation of these immune cells. The MIF signaling pathway involves CD74 interacting with CD44 a receptor that further propagates the signaling cascades essential for immune modulation and cell survival.
CD74 is linked to inflammatory and autoimmune conditions such as rheumatoid arthritis and systemic lupus erythematosus. Aberrant expression of CD74 influences the presentation of autoantigens and may contribute to the chronic inflammation seen in these disorders. Its connection to MIF which is a pro-inflammatory cytokine further implicates CD74 in these autoimmune scenarios by promoting persistent immune activation. Researchers are also exploring CD74 as a therapeutic target in certain cancers due to its influence on immune surveillance and tumor progression.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Overlay histogram showing Raji cells stained with ab9514 (red line). The cells were fixed with 80% methanol (5 min) and 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 (ab9514, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
Formalin-fixed, paraffin-embeded human tonsil tissue stained for CD74 using ab9514 at 1/50 dilution in immunohistochemical analysis. Antigen retrieval with citrate buffer pH 6.0.
All lanes: Western blot - Anti-CD74 antibody [LN2] (ab9514) at 5 µg/mL
All lanes: Human tonsil normal tissue lysate - total protein (ab29615) at 10 µg
All lanes: Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDa, 37 kDa
Flow cytometry overlay histogram showing wild-type Raji (green line) and CD74 knockout Raji cells (Human CD74 knockout Raji cell line ab273378) stained with ab9514 (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 (ab9514) (1x106 in 100μl at 1 μg/ml) for 30 min at 22°C.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) was used at 1/2000 for 30 min at 22°C.
Isotype control antibody was mouse IgG1κ (Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control ab170190) used at the same concentration and conditions as the primary antibody (wild-type Raji cells - black line; CD74 knockout Raji cells Human CD74 knockout Raji cell line 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 antibody can also be used in Raji cells fixed with 4% formaldehyde (10 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.
ab9514 staining CD74 in paraffin embedded Human tonsil tissue sections by Immunohistochemistry (IHC-P).
ab9514 staining CD74 in human liver tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with neutral buffered formalin and a heat mediated antigen retrieval step was performed using EDTA buffer pH 9.0. Samples were then blocked with 10% serum for 10 minutes at 20°C followed by incubation with the primary antibody at a 1/75 dilution for 30 minutes at 20°C. A HRP-conjugated rat anti-mouse/rabbit polyclonal was used undiluted as the secondary antibody.
Lanes 1 - 4: Merged signal (red and green). Green - ab9514 observed at 35 kDa. Red - loading control, Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.
ab9514 was shown to react with CD74 in western blot. The band observed in CD74 knockout cell line Human CD74 knockout Raji cell line ab273378 (knockout lysate Human CD74 knockout Raji cell 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 ab9514 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at 5 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-CD74 antibody [LN2] (ab9514) at 5 µg/mL
Lane 1: Wild-type Raji cell lysate at 30 µg
Lane 2: CD74 knockout Raji cell lysate at 30 µg
Lane 3: Jurkat cell lysate at 30 µg
Lane 4: HepG2 cell lysate at 30 µg
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 35 kDa
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