CD74 Antibody [Unconjugated]

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.

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 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.

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.

• Reactivity: Mu
• Applications: WB, Simple Western, IHC
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

CD74 Antibody [Unconjugated] Summary

• 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.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: CD74
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Simple Western 20 ug/mL
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CD74 Antibody [Unconjugated]

• CD74 antigen (invariant polypeptide of major histocompatibility complex, classII antigen-associated)
• CD74 antigen
• CD74 molecule, major histocompatibility complex, class II invariant chain
• CD74
• CLIP
• DHLAG
• DHLAGgamma chain of class II antigens
• HLA class II histocompatibility antigen gamma chain
• HLADG
• HLA-DR antigens-associated invariant chain
• HLA-DR-gamma
• Ia antigen-associated invariant chain
• Ia-associated invariant chain
• Ia-gamma
• Ii
• INVG34
• MHC HLA-DR gamma chain
• p33

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CD74 Antibody (AF7478)(7)

Publications using AF7478

• Qiu, Z;Khalife, J;Ethiraj, P;Jaafar, C;Lin, AP;Holder, KN;Ritter, JP;Chiou, L;Huelgas-Morales, G;Aslam, S;Zhang, Z;Liu, Z;Arya, S;Gupta, YK;Dahia, PLM;Aguiar, RCT; IRF8-mutant B cell lymphoma evades immunity through a CD74-dependent deregulation of antigen processing and presentation in MHCII complexes Science advances 2024-07-12 [PMID: 38996030] (Westen Blot)
• Gruel, R;Bijnens, B;Van Den Daele, J;Thys, S;Willems, R;Wuyts, D;Van Dam, D;Verstraelen, P;Verboven, R;Roels, J;Vandamme, N;Mancuso, R;Pita-Almenar, JD;De Vos, WH; S100A8-enriched microglia populate the brain of tau-seeded and accelerated aging mice Aging cell 2024-05-01 [PMID: 38403918] (Immunohistochemistry, Mouse)
• Qiu, Z;Khalife, J;Lin, AP;Ethiraj, P;Jaafar, C;Chiou, L;Huelgas-Morales, G;Aslam, S;Arya, S;Gupta, YK;Dahia, PLM;Aguiar, RCT; IRF8-mutant B cell lymphoma evades immunity through a CD74-dependent deregulation of antigen processing and presentation in MHC CII complexes bioRxiv : the preprint server for biology 2023-10-15 [PMID: 37873241] (Western Blot, Mouse)
• Ebert, S;Zang, L;Ismail, N;Otabil, M;Fröhlich, A;Egea, V;Ács, S;Hoeberg, M;Berres, ML;Weber, C;Moreira, JMA;Ries, C;Bernhagen, J;El Bounkari, O; Tissue Inhibitor of Metalloproteinases-1 Interacts with CD74 to Promote AKT Signaling, Monocyte Recruitment Responses, and Vascular Smooth Muscle Cell Proliferation Cells 2023-07-20 [PMID: 37508563] (Immunocytochemistry, Human)
• Z Feng, J Zhou, Y Liu, R Xia, Q Li, L Yan, Q Chen, X Chen, Y Jiang, G Chao, M Wang, G Zhou, Y Zhang, Y Wang, H Xia Epithelium- and endothelium-derived exosomes regulate the alveolar macrophages by targeting RGS1 mediated calcium signaling-dependent immune response Cell Death and Differentiation, 2021-03-22;0(0):. 2021-03-22 [PMID: 33753901] (ELISA Control, Mouse)
• L Farr, S Ghosh, N Jiang, K Watanabe, M Parlak, R Bucala, S Moonah CD74 signaling links inflammation to intestinal epithelial cell regeneration and promotes mucosal healing Cell Mol Gastroenterol Hepatol, 2020-01-28;0(0):. 2020-01-28 [PMID: 32004754] (IHC, Mouse)
• Akinobu Ochi, Dong Chen, Wibke Schulte, Lin Leng, Nickolas Moeckel, Marta Piecychna et al. MIF-2/D-DT enhances proximal tubular cell regeneration through SLPI- and ATF4-dependent mechanisms American Journal of Physiology-Renal Physiology 9/1/2017 [PMID: 28539339]

Bioinformatics

• Gene Symbol: CD74
• Uniprot:
  • NP_034675()