Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Simple Western

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry, ELISA Control, Westen Blot

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all CD74 Primary Antibodies »

Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant mouse CD74
Gln56-Leu215
Accession # NP_034675

Specificity

Detects mouse CD74 in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross-reactivity with recombinant human CD74 is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Mouse CD74 Antibody

Detection of Mouse CD74 antibody by Western Blot.

Detection of Mouse CD74 by Western Blot.

Western blot shows lysates of A20 mouse B cell lymphoma cell line and mouse spleen tissue. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Mouse CD74 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7478) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for CD74 at approximately 31-34 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

CD74 antibody in Mouse Liver by Immunohistochemistry (IHC-Fr).

CD74 in Mouse Liver.

CD74 was detected in perfusion fixed frozen sections of mouse liver using Sheep Anti-Mouse CD74 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7478) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; CTS019) and counter-stained with hematoxylin (blue). Specific staining was localized to stellate cells. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Mouse CD74 by Simple WesternTM.

Simple Western lane view shows lysates of Mouse Spleen, loaded at 0.2 mg/mL. A specific band was detected for CD74 at approximately 47 kDa (as indicated) using 20 µg/mL of Sheep Anti-Mouse CD74 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7478) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Detection of CD74 by Immunocytochemistry/ Immunofluorescence

Detection of CD74 by Immunocytochemistry/ Immunofluorescence

Validation of the interaction between TIMP-1 and CD74 by immunocytochemistry and immunoprecipitation analysis. (a) Colocalization of TIMP-1 and CD74 in THP-1 monocytes. Fluorescence microscopy analysis costained with anti-TIMP-1/Star635 secondary antibody (red), anti-CD74/Star488 secondary antibody (green), and DAPI (blue, DNA labeling in nuclei). The merged image shows the overlay (yellow) of the two channels demonstrating TIMP-1 colocalization with CD74. Arrows indicate membrane regions of substantial colocalization. Scale bar: 10 µm. (b,c) Immunoprecipitation of TIMP-1 with cell surface CD74. Semi-endogenous pull-down assay using protein cell lysate from HEK29 cells transfected with FLAG-tagged CD74 and recombinant TIMP-1 analyzed by Western blotting with anti-CD74 (b) and anti-TIMP-1 (c) antibodies. (d,e) TIMP-1 interacts with the soluble CD74 ectodomain fragment (sCD74). Immunoprecipitation of His-tagged sCD74 and recombinant TIMP-1 and the analysis of precipitated proteins by Western blotting using anti-His (d) and anti-TIMP-1 (e) antibodies. C: control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37508563), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CD74 by Immunocytochemistry/ Immunofluorescence

Detection of CD74 by Immunocytochemistry/ Immunofluorescence

Validation of the interaction between TIMP-1 and CD74 by immunocytochemistry and immunoprecipitation analysis. (a) Colocalization of TIMP-1 and CD74 in THP-1 monocytes. Fluorescence microscopy analysis costained with anti-TIMP-1/Star635 secondary antibody (red), anti-CD74/Star488 secondary antibody (green), and DAPI (blue, DNA labeling in nuclei). The merged image shows the overlay (yellow) of the two channels demonstrating TIMP-1 colocalization with CD74. Arrows indicate membrane regions of substantial colocalization. Scale bar: 10 µm. (b,c) Immunoprecipitation of TIMP-1 with cell surface CD74. Semi-endogenous pull-down assay using protein cell lysate from HEK29 cells transfected with FLAG-tagged CD74 and recombinant TIMP-1 analyzed by Western blotting with anti-CD74 (b) and anti-TIMP-1 (c) antibodies. (d,e) TIMP-1 interacts with the soluble CD74 ectodomain fragment (sCD74). Immunoprecipitation of His-tagged sCD74 and recombinant TIMP-1 and the analysis of precipitated proteins by Western blotting using anti-His (d) and anti-TIMP-1 (e) antibodies. C: control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37508563), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse CD74 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of mouse liver

Simple Western

20 µg/mL
Sample: Mouse Spleen

Western Blot

2 µg/mL
Sample: A20 mouse B cell lymphoma cell line and mouse spleen tissue

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CD74

CD74 (also known as H2 class II histocompatibility antigen gamma chain and Ia antigen-associated invariant chain/Ii) is 31-41 kDa glycoprotein, member of no known protein family. It is expressed by multiple cell types, including dendritic cells, macrophages, immature and mature B cells, type II greater alveolar epithelial cells and colonic epithelium. CD74 is best known as an ER-embedded trimeric chaperone that directs MHC class II dimers through the endosome and into the cell membrane. When embedded in the endosome, it undergoes cleavage that both facilitates MHC dimer transit, and generates a cytosolic signaling fragment. It also appears on the cell surface where, in conjunction with CD44 (and possibly c-met), it binds MIF, inducing either cell proliferation, cell survival,or cytokine release. Mouse CD74 is a 279 amino acid (aa) type II transmembrane protein (SwissProt P04441). It possesses a 30 aa cytoplasmic region plus a 224 aa luminal/extracellular domain (aa 56‑279) that contains one thyroglobulin type-1 repeat (aa 193-254). CD74 is known to be phosphorylated on Ser9, and occasionally undergo glycanation with chrondroitin at Ser265 (on monocytes and B cells). There are multiple potential splice forms. The isoform used for antibody formation by R&D Systems, Inc. exhibits a deletion of aa 192-255 (GenBank NP_034675). Two other splice forms show either the same deletion just described coupled to a four aa substitution for aa 277-279, or a 10 aa substitution for aa 131-279. Over aa 56-215 of the NP_034675 isoform, mouse CD74 shares 86% and 69% aa sequence identity with rat and human CD74, respectively.

Alternate Names

CD74, CLIP, DHLAG, HLADG, Ia-gamma, INVG34

Entrez Gene IDs

972 (Human); 16149 (Mouse); 25599 (Rat)

Gene Symbol

CD74

UniProt

NP_034675

Additional CD74 Products

Product Documents for Mouse CD74 Antibody

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Product Specific Notices for Mouse CD74 Antibody

For research use only

Citations for Mouse CD74 Antibody

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Protocols

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