ET1702-51

CD74 Recombinant Rabbit Monoclonal Antibody [JF40-10]

cat.: ET1702-51

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JF40-10

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 34 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human CD74 aa 21-64 / 296.
Positive control:	Raji cell lysate, Jurkat cell lysate, Raji, human spleen tissue, human lymph nodes tissue, human tonsil tissue.
Subcellular location:	Endoplasmic reticulum membrane, trans-Golgi network, cell membrane, lysosome, endosome.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:1,000-1:5,000 1:50-1:200 1:50-1:500 1:500 1:1,000
Uniprot #:	SwissProt: P04233 Human
Alternative names:	CD 74 CD74 CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated) CD74 antigen CD74 molecule CD74 molecule, major histocompatibility complex, class II invariant chain CLIP DHLAG Gamma chain of class II antigens HG2A_HUMAN HLA class II histocompatibility antigen gamma chain HLA DR antigens associated invariant chain HLA DR gamma HLA-DR antigens-associated invariant chain HLA-DR-gamma HLADG HLADR antigens associated invariant chain Ia antigen associated invariant chain Ia antigen-associated invariant chain Ia associated invariant chain Ia gamma Ii Invariant polypeptide of major histocompatibility complex class II antigen associated la-gamma Major histocompatibility complex class II invariant chain MHC HLA DR gamma chain MHC HLA-DR gamma chain p33 p35 Protein 41

Images

Fig1: Western blot analysis of CD74 on different lysates with Rabbit anti-CD74 antibody (ET1702-51) at 1/1,000 dilution.

Lane 1: Raji cell lysate
Lane 2: Jurkat cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 34 kDa
Observed band size: 34 kDa

Exposure time: Lane 1: 59 seconds; Lane 2: 3 minutes; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-51) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of Raji cells labeling CD74 with Rabbit anti-CD74 antibody (ET1702-51) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD74 antibody (ET1702-51) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD74 antibody (ET1702-51) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1702-51) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig4: Immunohistochemical analysis of paraffin-embedded human lymph nodes tissue with Rabbit anti-CD74 antibody (ET1702-51) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1702-51) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD74 antibody. Counter stained with hematoxylin.
	Fig6: Flow cytometric analysis of Raji cells labeling CD74. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-51, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.