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Rabbit Recombinant Monoclonal CD74 antibody. Suitable for Flow Cyt, WB, IHC-P and reacts with Human samples. Immunogen corresponding to Cell preparation containing CD74 protein.


Images

Flow Cytometry - Anti-CD74 antibody [CLIP/3127R] (AB270265), expandable thumbnail
  • Western blot - Anti-CD74 antibody [CLIP/3127R] (AB270265), expandable thumbnail
  • Flow Cytometry - Anti-CD74 antibody [CLIP/3127R] (AB270265), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [CLIP/3127R] (AB270265), expandable thumbnail
  • Western blot - Anti-CD74 antibody [CLIP/3127R] (AB270265), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • Cell preparation containing CD74 protein. The exact immunogen used to generate this antibody is proprietary information. Database link P04233

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
Flow CytWBIHC-P
Human
Tested
Tested
Tested
Mouse
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Baboon
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

1.00000-2.00000 µg for 106 Cells

Notes

-

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Mouse

Dilution info

-

Notes

-

Species

Baboon

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1.00000-2.00000 µg/mL

Notes

-

Not recommended
Not recommended

Species

Rat, Mouse, Baboon

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1.00000-2.00000 µg/mL

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species

Rat

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Mouse

Dilution info

-

Notes

-

Species

Baboon

Dilution info

-

Notes

-

Associated Products

Select an associated product type

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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 Recombinant Monoclonal CD74 antibody. Suitable for Flow Cyt, WB, IHC-P and reacts with Human samples. Immunogen corresponding to Cell preparation containing CD74 protein.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • Cell preparation containing CD74 protein. The exact immunogen used to generate this antibody is proprietary information. Database link P04233
Clone number

CLIP/3127R

Purification technique

Affinity purification Protein A/G

Concentration
Loading...
Purification notes

Purified from Bioreactor Concentrate by Protein A/G.

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

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.

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5 product images

  • Flow Cytometry - Anti-CD74 antibody [CLIP/3127R] (ab270265), expandable thumbnail

    Flow Cytometry - Anti-CD74 antibody [CLIP/3127R] (ab270265)

    Flow cytometry overlay histogram showing wild-type Raji (green line) and CD74 knockout Raji cells (Human CD74 knockout Raji cell line ab273378) stained with ab270265 (red line). The cells were 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 (ab270265) (1x106 in 100μl at 1 μg/ml) for 30 min at 4°C.

    The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) was used at 1/2000 for 30 min at 4°C.

    Isotype control antibody was Rabbit IgG (monoclonal) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) 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.

  • Western blot - Anti-CD74 antibody [CLIP/3127R] (ab270265), expandable thumbnail

    Western blot - Anti-CD74 antibody [CLIP/3127R] (ab270265)

    Lanes 1 - 4: Merged signal (red and green). Green - ab270265 observed at 35 kDa. Red - loading control, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab270265 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 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab270265 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 [CLIP/3127R] (ab270265) 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

  • Flow Cytometry - Anti-CD74 antibody [CLIP/3127R] (ab270265), expandable thumbnail

    Flow Cytometry - Anti-CD74 antibody [CLIP/3127R] (ab270265)

    Flow cytometric analysis of Raji cells labeling CD74 using ab270265 at 2 μg/106 cells (Blue), followed by goat anti-mouse IgG-CF488; isotype control (Red).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [CLIP/3127R] (ab270265), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [CLIP/3127R] (ab270265)

    Formalin-fixed, paraffin-embedded human tonsil stained for CD74 using ab270265 at 2 μg/ml in imunnohistochemical analysis.

  • Western blot - Anti-CD74 antibody [CLIP/3127R] (ab270265), expandable thumbnail

    Western blot - Anti-CD74 antibody [CLIP/3127R] (ab270265)

    All lanes: Western blot - Anti-CD74 antibody [CLIP/3127R] (ab270265) at 2 µg/mL

    All lanes: HEK-293T cell lysate

    Predicted band size: 34 kDa

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Product protocols

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