Revision 1

#70167

Store at -20C

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, IF-F, IF-IC

Reactivity:
M

Sensitivity:
Endogenous

MW (kDa):
31

Source/Isotype:
Rabbit IgG

UniProt ID:
#P04441

Entrez-Gene Id:
16149

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Frozen) 1:400 - 1:800
Immunofluorescence (Immunocytochemistry) 1:800 - 1:1600

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

CD74 (F6K1S) Rabbit Monoclonal Antibody recognizes endogenous levels of total CD74 protein. This antibody detects multiple bands that are consistent with multimerization of CD74.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gln272 of mouse CD74 protein.

Background

CD74, which is also known as the MHC Class II-associated invariant chain (Ii), is a type II transmembrane glycoprotein that plays a critical role in the antigen presentation process as a chaperone of MHC Class II proteins. It is expressed at high levels on B cells and to a lesser extent on numerous antigen presenting cell (APC) types including dendritic cells, Langerhans cells, monocytes, and macrophages as well as non-traditional APCs such as epithelial cells (1,2). CD74 was initially identified for its ability to regulate the folding and intracellular trafficking of newly synthesized MHC Class II molecules. Following expression, CD74 self-assembles as a trimer that serves as a scaffold for the assembly of MHC Class II molecules. Through this interaction, CD74 blocks the peptide binding cleft of MHC Class II molecules and prevents their premature association with endogenous polypeptides (3). Binding to CD74 also facilitates the translocation of MHC Class II molecules from the endoplasmic reticulum to the endocytic compartments during antigen presentation (4). In addition to its role as an MHC Class II chaperone, CD74 is also the receptor for macrophage migration-inhibitory factor (MIF). Binding to CD74 and its co-receptor, CD44, has been shown to induce the activation of the NFkB and ERK pathways to promote cell proliferation and survival signals (5,6). Recent studies have identified CXCR2 and CXCR4 as co-receptors for CD74 where MIF binding to CD74 complexes contributes to MIF-mediated monocyte chemotaxis and the induction of Akt signaling, respectively (7,8). Increased CD74 surface expression has been reported under inflammatory conditions and in certain types of cancer cells implying a potential role in tumorigenesis (9).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

M: Mouse

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

使用に関する制限

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Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from A20 and MEFs, Wt cells, and mouse spleen tissue using CD74 (F6K1S) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Negative expression of CD74 protein in MEFs, Wt cells is consistent with the predicted expression pattern.
Western Blotting Image 1: CD74 (F6K1S) Rabbit Monoclonal Antibody
Western blot analysis of extracts from mouse spleen tissue, untreated (-) or treated with peptide N-glycosidase F (PNGase F; +), using CD74 (F6K1S) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 2: CD74 (F6K1S) Rabbit Monoclonal Antibody
Immunoprecipitation of CD74 protein from mouse spleen tissue extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is CD74 (F6K1S) Rabbit mAb. Western blot analysis was performed using CD74 (F6K1S) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation Image 1: CD74 (F6K1S) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of fixed frozen subiculum from an amyloid mouse model of Alzheimer's disease using CD74 (F6K1S) Rabbit mAb (green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to co-labeling with β-Amyloid (D54D2) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) #35363 (red), ASC/TMS1 (D2W8U) Rabbit mAb (Alexa Fluor® 647 Conjugate) #23640 (white), and ProLong Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 1: CD74 (F6K1S) Rabbit Monoclonal Antibody
Confocal immunofluorescent analysis of barrier associated macrophages from fixed frozen mouse brain using CD74 (F6K1S) Rabbit mAb (green), MHC Class II (I-A/I-E) (M5/114.15.2) Rat mAb #86285 (red), and ProLong Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 2: CD74 (F6K1S) Rabbit Monoclonal Antibody
Confocal immunofluorescent analysis of fixed frozen mouse spleen using CD74 (F6K1S) Rabbit mAb (green) and co-labeled with MHC Class II (I-A/I-E) (M5/114.15.2) Rat mAb #86285 (red) and ProLong Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 3: CD74 (F6K1S) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of fixed frozen mouse colon using CD74 (F6K1S) Rabbit mAb (green), MHC Class II (I-A/I-E) (M5/114.15.2) Rat mAb #86285 (red), and ProLong Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 4: CD74 (F6K1S) Rabbit Monoclonal Antibody
Confocal immunofluorescent analysis of RAW 264.7 cells (left, positive) and MEF cells (right, negative) using CD74 (F6K1S) Rabbit mAb (green), DyLight 554 Phalloidin #13054 (red), and DAPI #4083 (blue).
Immunofluorescence Image 1: CD74 (F6K1S) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.