WB
Western blot analysis of CD74 on different lysates with Rabbit anti-CD74 antibody at 1/1,000 dilution. Lane 1: Raji cell lysate, Lane 2: Jurkat cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: Lane 1: 59 seconds; Lane 2: 3 minutes; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD74 antibody 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 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.IHC
Immunohistochemical analysis of paraffin-embedded human lymph nodes tissue with Rabbit anti-CD74 antibody 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 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.ICC/IF
Immunocytochemistry analysis of Raji cells labeling CD74 with Rabbit anti-CD74 antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of Raji cells labeling CD74. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IF-P
Species: Human, Tissue: Spleen, Sample: Paraffin-embedded section, Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃. Wash buffer: 1× TBST, Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. Primary antibody: 1/200, overnight at 4℃. Secondary antibody: Goat Anti-Rabbit IgG (488), 1.5 hours at room temperature.| Product Name | CD74 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | The human histocompatibility leukocyte antigen (HLA) class II-associated invariant chain is composed of at least four polypeptides. One of these polypeptide chains is expressed as a membrane-bound subunit and has been designated CD74. The loading of peptide onto the class II MHC protein (MHC II) appears to be regulated by CD74, which associates with MHC II during its migration to the endosomal compartment, where class II binds pep-tide. CD74 is expressed by cells of both T lymphocyte and B lymphocyte lineages. In fact, CD74 is broadly expressed in normal B lymphocytes, regardless of their histocompatibility leukocyte antigen (HLA) phenotype, while a subset of peripheral T lymphocytes that are MHC II negative do not express CD74. |
| Immunogen | Synthetic peptide within Human CD74 aa 21-64 / 296. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIF-PIHCWB |
| WB:1:1000-1:5000 IHC:1:500 ICC/IF:1:50-1:200 FC:1:1000 IF-P:1:200 |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | CD 74 antibody
CD74 antibody
CD74 antigen (invariant polypeptide of major histocompatibility complex class II antigen-associated) antibody CD74 antigen antibody CD74 molecule antibody CD74 molecule major histocompatibility complex class II invariant chain antibody CLIP antibody DHLAG antibody Gamma chain of class II antigens antibody HG2A_HUMAN antibody HLA class II histocompatibility antigen gamma chain antibody HLA DR antigens associated invariant chain antibody HLA DR gamma antibody HLA-DR antigens-associated invariant chain antibody HLA-DR-gamma antibody HLADG antibody HLADR antigens associated invariant chain antibody Ia antigen associated invariant chain antibody Ia antigen-associated invariant chain antibody Ia associated invariant chain antibody Ia gamma antibody Ii antibody Invariant polypeptide of major histocompatibility complex class II antigen associated antibody la-gamma antibody Major histocompatibility complex class II invariant chain antibody MHC HLA DR gamma chain antibody MHC HLA-DR gamma chain antibody p33 antibody p35 antibody Protein 41 antibody |
|---|---|
| Molecular Weight(MW) | 34 kDa |
| Cellular Localization | Endoplasmic reticulum membrane, trans-Golgi network, cell membrane, lysosome, endosome. |
| SwissProt ID | P04233 |
|---|
WB
Western blot analysis of CD74 on different lysates with Rabbit anti-CD74 antibody at 1/1,000 dilution. Lane 1: Raji cell lysate, Lane 2: Jurkat cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: Lane 1: 59 seconds; Lane 2: 3 minutes; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD74 antibody 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 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.
IHC
Immunohistochemical analysis of paraffin-embedded human lymph nodes tissue with Rabbit anti-CD74 antibody 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 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.
ICC/IF
Immunocytochemistry analysis of Raji cells labeling CD74 with Rabbit anti-CD74 antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of Raji cells labeling CD74. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IF-P
Species: Human, Tissue: Spleen, Sample: Paraffin-embedded section, Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃. Wash buffer: 1× TBST, Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. Primary antibody: 1/200, overnight at 4℃. Secondary antibody: Goat Anti-Rabbit IgG (488), 1.5 hours at room temperature.| Application Notes | WB:1:1000-1:5000 IHC:1:500 ICC/IF:1:50-1:200 FC:1:1000 IF-P:1:200 |
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| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
