Anti-CD74 Antibody (56451)

Host Quantity Applications Species Reactivity Data Sheet  
Mouse 100ug WB,IHC,ICC/IF,IP,FCM,FACS Human
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Overview
Product Name Anti-CD74 Antibody (56451)
Description Anti-CD74 Mouse Monoclonal Antibody
Target CD74
Species Reactivity Human
Applications WB,IHC,ICC/IF,IP,FCM,FACS
Host Mouse
Clonality Monoclonal
Clone ID PIN1
Isotype IgG1
Immunogen Synthetic peptide corresponding to the human CD74 invariant chain (aa 12-28) conjugated to KLH.
Properties
Form Liquid
Concentration Lot Specific
Formulation PBS, pH 7.4.
Buffer Formulation Phosphate Buffered Saline
Buffer pH pH 7.4
Format Purified
Purification Purified by Protein G affinity chromatography
Specificity Information
Specificity This antibody recognizes human CD74.
Target Name HLA class II histocompatibility antigen γ chain
Target ID CD74
Uniprot ID P04233
Alternative Names HLA-DR antigens-associated invariant chain, Ia antigen-associated invariant chain, Ii, CD antigen CD74 [Cleaved into: Class-II-associated invariant chain peptide
Gene Name CD74
Sequence Location Cell membrane, Endoplasmic reticulum membrane. Golgi apparatus, trans-Golgi network. Endosome. Lysosome. Note=Transits through a number of intracellular compartments in the endocytic pathway. It can either undergo proteolysis or reach the cell membrane.; [Isoform p41]: Late endosome, Lysosome
Biological 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 peptide]: Binds 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. {PubMed:1448172}.; [Isoform p41]: Stabilizes 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). {UniProtKB:P04441, PubMed:32855215}.
Research Areas Neuroscience
Background CD74 is a non-polymorphic type II integral membrane protein. It has a short N-terminal cytoplasmic tail of 28 amino acids followed by a single 24-aa transmembrane region and an approximately 150-aa lumenal domain. CD74 functions mainly as an MHC class II chaperone which promotes ER exit of MHC class II molecules by directing them to endocytic compartments, preventing peptide binding in the ER, and contributing to peptide “editing†in the MHC class II compartment. CD74 has recently been shown to have a role as an accessory signaling molecule because of its high- affinity binding to the pro-inflammatory cytokine macrophage migration inhibitory factor (MIF).
Application Images
Description Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-CD74 Monoclonal Antibody, Clone PIN 1.1 (56451). Tissue: HaCaT cells. Species: Human. Fixation: Cold 100% methanol for 10 minutes at -20°C. Primary Antibody: Mouse Anti-CD74 Monoclonal Antibody (56451) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Cytoplasmic Staining.
Description Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-CD74 Monoclonal Antibody, Clone PIN 1.1 (56451). Tissue: Cervical cancer cell line (HeLa). Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Mouse Anti-CD74 Monoclonal Antibody (56451) at 1:100 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus. Endosome. Lysosome. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-CD74 Antibody. (C) Composite.
Description Immunohistochemistry analysis using Mouse Anti-CD74 Monoclonal Antibody, Clone PIN 1.1 (56451). Tissue: backskin. Species: Mouse. Fixation: Bouin's Fixative and paraffin-embedded. Primary Antibody: Mouse Anti-CD74 Monoclonal Antibody (56451) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Beautiful basal to suprabasal staining in epidermis, dermis, hair follicles and muscle.
Description Western Blot analysis of Human N87 cell lysates showing detection of CD74 protein using Mouse Anti-CD74 Monoclonal Antibody, Clone PIN 1.1 (56451). Primary Antibody: Mouse Anti-CD74 Monoclonal Antibody (56451) at 1:1000. Lysates treated with macrophage inhibitory factor (MIF). Courtesy of: Victor E. Reyes, University of Texas Medical Branch, USA.
Additional Information
Handling
Storage This antibody is stable for at least one (1) year at -20°C.
Dilution Instructions Dilute in PBS or medium that is identical to that used in the assay system.
Application Instructions Immunoblotting: use at 1ug/mL. A doublet band of ~33-35 kDa is detected.

Immunoprecipitation: use at 12ug/mL.

Immunocytochemistry: working concentration is sample dependent.Flow cytometry: working concentration is sample dependent.

These are recommended concentrations. User should determine optimal concentrations for their application.

Positive control: PALA cell lysate.
References & Data Sheet