CD74 Recombinant Rabbit mAb (S-4524-58)

Catalog Number: S0B60489 Application: WB, IHC-P, ICC, FCM Reactivity: Hu Conjugation: Unconjugated Brand: Starter

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$100 USD

Size: 25μl 100μl

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Product Details

Product Specification

Host	Rabbit
Antigen	CD74
Synonyms	HLA class II histocompatibility antigen gamma chain; HLA-DR antigens-associated invariant chain; Ia antigen-associated invariant chain (Ii); DHLAG
Immunogen	Recombinant Protein
Location	Secreted, Cell membrane
Accession	P04233
Clone Number	S-4524-58
Antibody Type	Recombinant mAb
Isotype	IgG
Application	WB, IHC-P, ICC, FCM
Reactivity	Hu
Positive Sample	Raji, Daudi, Ramos, U266B1, human peripheral blood lymphocytes
Purification	Protein A
Concentration	0.5 mg/ml
Conjugation	Unconjugated
Physical Appearance	Liquid
Storage Buffer	PBS, 40% Glycerol, 0.05% BSA, 0.02% sodium azide
Stability & Storage	12 months from date of receipt / reconstitution, -20 °C as supplied

Dilution

application	dilution	species
WB	1:5000-1:20000	Hu
IHC-P	1:500-1:2000	Hu
ICC	1:500	Hu
FCM	1:50	Hu

Background

CD74, also known as the MHC class II-associated invariant chain (Ii chain), is a functionally versatile and critically important molecule in the immune system. Its most canonical function is to serve as a "chaperone" protein for MHC class II molecules in antigen-presenting cells, where it stabilizes the peptide-binding groove of MHC class II molecules, prevents premature binding of endogenous peptides, and directs their trafficking from the endoplasmic reticulum to endosomes/lysosomes, thereby ensuring efficient presentation of exogenous antigens to initiate specific immune responses. However, the functions of CD74 extend far beyond this: it also acts as a high-affinity cell surface receptor for the pro-inflammatory cytokine macrophage migration inhibitory factor (MIF). Upon binding of MIF to CD74—often in cooperation with CD44—multiple signaling pathways including NF-κB and PI3K-Akt are activated, subsequently regulating cell proliferation, survival, and inflammatory responses. This dual identity in both antigen presentation and inflammatory signaling casts CD74 as a "double-edged sword" in various diseases—it participates in B-cell development and the maintenance of immune homeostasis on one hand, while on the other, it is found to be aberrantly overexpressed in autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus, as well as in numerous hematological malignancies and solid tumors, where it promotes disease progression through its survival signaling pathways. Consequently, CD74-targeted therapeutic strategies (such as the humanized antibody Milatuzumab) and explorations of its utility as a diagnostic and prognostic biomarker in cancer and inflammatory diseases have become hot topics in current research.

Picture

Western Blot

WB result of CD74 Recombinant Rabbit mAb
Primary antibody: CD74 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: Raji whole cell lysate 20 µg
Lane 2: Daudi whole cell lysate 20 µg
Lane 3: Ramos whole cell lysate 20 µg
Lane 4: U266B1 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 26 kDa
Observed MW: 30-38 kDa

FC

Flow cytometric analysis of human peripheral blood lymphocytes labeled with Anti-Human CD74 antibody at 1/50 dilution (1 μg) / (right panel) compared with a Rabbit IgG Isotype Control / (left panel). Goat Anti- Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody. Flow cytometry and data analysis were performed using Agilent NovoCyte Quanteon and FlowJo™ software.

Immunohistochemistry

IHC shows positive staining in paraffin-embedded human liver. Anti-CD74 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human lung. Anti-CD74 antibody was used at 1/500 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human spleen. Anti-CD74 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human tonsil. Anti-CD74 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human diffuse large B-cell lymphoma. Anti-CD74 antibody was used at 1/500 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human ovarian cancer. Anti-CD74 antibody was used at 1/500 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

Immunocytochemistry

ICC shows positive staining in Raji cells (top panel) and negative staining in Jurkat cells (below panel). Anti-CD74 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibod at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).