Rabbit Polyclonal CD74 antibody. Suitable for IHC-P, IP, WB, ICC/IF and reacts with Human samples. Cited in 6 publications.
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- Cell regeneration (London, England) 12:272023Cross-species single-cell transcriptomic analysis of animal gastric antrum reveals intense porcine mucosal immunity.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXiaodan Wang,Fan Hong,Haonan Li,Yalong Wang,Mengxian Zhang,Shibo Lin,Hui Liang,Hongwen Zhou,Yuan Liu,Ye-Guang ChenPubMed 37525021
- Acta neuropathologica communications 11:692023Microglial CD68 and L-ferritin upregulation in response to phosphorylated-TDP-43 pathology in the amyotrophic lateral sclerosis brain.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMolly E V Swanson,Miran Mrkela,Helen C Murray,Maize C Cao,Clinton Turner,Maurice A Curtis,Richard L M Faull,Adam K Walker,Emma L ScotterPubMed 37118836
- Communications biology 4:5632021A follicular regulatory Innate Lymphoid Cell population impairs interactions between germinal center Tfh and B cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMargaret H O'Connor,Roshell Muir,Marita Chakhtoura,Michael Fang,Eirini Moysi,Susan Moir,Alison J Carey,Alyssa Terk,Carmen N Nichols,Talibah Metcalf,Constantinos Petrovas,Mark J Cameron,Virginie Tardif,Elias K HaddadPubMed 33980982
- Scientific reports 10:116932020Quantitative immunohistochemical analysis of myeloid cell marker expression in human cortex captures microglia heterogeneity with anatomical context.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMolly E V Swanson et. al.PubMed 32678124
- Methods in molecular biology (Clifton, N.J.) 2080:123-1342019Concurrent Immunohistochemical Localization and Western Blot Analysis of the MIF Receptor, CD74, in Formalin-Fixed, Paraffin-Embedded Tissue.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAmanda Graham,Warren B NothnickPubMed 31745876
- Reproductive sciences (Thousand Oaks, Calif.) 25:1557-15662018Macrophage Migration Inhibitory Factor Receptor, CD74, is Overexpressed in Human and Baboon ( Papio Anubis) Endometriotic Lesions and Modulates Endometriotic Epithelial Cell Survival and Interleukin 8 Expression.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWarren B Nothnick,Tommaso Falcone,Mark R Olson,Asgerally T Fazleabas,Ossama W Tawfik,Amanda GrahamPubMed 29592775
Key facts
- Isotype
IgG
- Host species
Rabbit
- Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA- Form
Liquid
- Clonality
Polyclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data
Select an application| IHC-P | IP | WB | ICC/IF | |
|---|---|---|---|---|
| Human | Tested | Tested | Tested | Tested |
| Primates | Predicted | Predicted | Predicted | Predicted |
IHC-P
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1/80 - 1/250 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Primates | Dilution info - | Notes - |
IP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 5 µg/mL | Notes IP image from Phase V. Lot JLD 04.06.2013 |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Primates | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Primates | Dilution info - | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Primates | Dilution info - | Notes - |
Associated Products
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Target data
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.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).
Alternative names
DHLAG, CD74, DHLAG, HLA class II histocompatibility antigen gamma chain, HLA-DR antigens-associated invariant chain, Ia antigen-associated invariant chain, Ii
Recommended products
Rabbit Polyclonal CD74 antibody. Suitable for IHC-P, IP, WB, ICC/IF and reacts with Human samples. Cited in 6 publications.
Key facts
- Isotype
IgG
- Host species
Rabbit
- Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA- Form
Liquid
- Clonality
Polyclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Purification technique
Affinity purification Immunogen
- Concentration
Storage
- Shipped at conditions
Blue Ice
- Appropriate short-term storage duration
1-2 weeks
- Appropriate short-term storage conditions
+4°C
- Appropriate long-term storage conditions
-20°C
- Aliquoting information
Upon delivery aliquot
- Storage information
Avoid freeze / thaw cycle
Notes
Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.
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Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Biological function summary
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.
- Pathways
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.
- Associated diseases and disorders
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.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
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.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
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8 product images
This image is courtesy of a customer review submitted by Antibody Solutions Ltd.Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody (ab64772)
ab64772 (1/250) staining CD74 in paraffin-embedded Human tonsil tissue. Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval. The secondary antibody was goat anti rabbit conjugated to HRP. For further experimental details please refer to abreview.
Immunoprecipitation - Anti-CD74 antibody (ab64772)
CD74 was immunoprecipitated using 0.5mg Raji whole cell extract, 5μg of Rabbit polyclonal to CD74 and 50μl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Raji whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40μl SDS loading buffer and incubated for 10min at 70°C; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab64772.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) ab99697).
Band: 34kDa; CD74
All lanes: Immunoprecipitation - Anti-CD74 antibody (ab64772)
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 34 kDa
Exposure time: 12min
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody (ab64772)
ab64772 (1:160) staining CD74 in paraffin-embedded human tonsil (left panel) using an automated system (Ventana Discovery). Right-hand panel shows negative control (no primary antibody).
Using this protocol there is strong membrane staining of activated B cells in the germinal centres and B cells of the mantle zone of the follicles plus scattered cells of the interfollicular areas (paracortical B cells).Sections were rehydrated and antigen retrieved in CC1 Cell Conditioning Buffer using Ventana Mild Retrieval programme. Slides were blocked in 3% H2O2 / 4 min / 37°C and incubated with ab64772 (1:160 dilution / 2 hours / 37°C). Sections then blocked (4mins / 37°C) and incubated with Dako swine anti-rabbit antibody (1:50, 28 min / 37°C). Staining was amplified and detected by incubation with Ventana Streptavidin ABC system (16 min / 37°C) and Ventana DAB map reagent (8 min / 37°C). Slides were counterstained with Haematox
Western blot - Anti-CD74 antibody (ab64772)
Lanes 1 - 4: Merged signal (red and green). Green - ab64772 observed at 35 kDa. Red - loading control, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab64772 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 ab64772 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-CD74 antibody (ab64772) at 1 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
Western blot - Anti-CD74 antibody (ab64772)
All lanes: Western blot - Anti-CD74 antibody (ab64772) at 1 µg/mL
All lanes: Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate at 10 µg
SecondaryAll lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Exposure time: 2min
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody (ab64772)
ab64772 (1:80) staining CD74 in paraffin-embedded human lymph node (left panel) using an automated system (Ventana Discovery). Right-hand panel shows negative control (no primary antibody).
Using this protocol there is strong membrane staining of activated B cells in the germinal centres and B cells of the mantle zone of the follicles plus scattered cells of the interfollicular areas (paracortical B cells).Sections were rehydrated and antigen retrieved in CC1 Cell Conditioning Buffer using Ventana Mild Retrieval programme. Slides were blocked in 3% H2O2 / 4 min / 37°C and incubated with ab64772 (1:80 dilution / 1 hour / 37°C). Sections then blocked (3mins / 37°C) and incubated with Dako swine anti-rabbit antibody (1:50, 28 min / 37°C). Staining was amplified and detected by incubation with Ventana Streptavidin ABC system (16 min / 37°C) and Ventana DAB map reagent (8 min / 37°C). Slides were counterstained with Haematoxyli
Immunocytochemistry/ Immunofluorescence - Anti-CD74 antibody (ab64772)
ICC/IF image of ab64772 stained RAW246.7 cells. The cells were 4% paraformaldehyde fixed (10 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 (ab64772, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899, DyLight® 488 goat anti-rabbit 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.
Western blot - Anti-CD74 antibody (ab64772)
Lanes 1 - 4: Merged signal (red and green). Green - ab64772 observed at 35 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab64772 was shown to react with CD74 in wild-type Raji cells in Western blot with loss of signal observed in CD74 knockout cell line Human CD74 knockout Raji cell line ab273876 (knockout cell lysate Human CD74 knockout Raji cell lysate ab273830). Wild-type Raji and CD74 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab64772 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-CD74 antibody (ab64772) at 1 µg/mL
Lane 1: Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 2: CD74 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 3: Daudi (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 4: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 35 kDa
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