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CD74 Recombinant Mouse Monoclonal Antibody [PDM0-22]

Usd: 350

Specification

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Catalog# HA601117

CD74 Recombinant Mouse Monoclonal Antibody [PDM0-22]

Application

  • WB

  • IHC-P

  • IF-Cell

  • IF-Tissue

Reactivity

  • Human

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

CD74 Recombinant Mouse Monoclonal Antibody [PDM0-22]

Antibody Type

Recombinant Mouse Monoclonal Antibody

Immunogen

Recombinant protein within Human CD74 73-296 (Extracellular).

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Cell, IF-Tissue

Molecular Weight

Predicted band size: 33 kDa

Positive Control

Daudi cell lysate, Raji cell lysate, LOVO, human B-cell lymphoma tissue, human tonsil tissue, human spleen tissue, human prostate cancer tissue.

Conjugation

unconjugated

Clone Number

PDM0-22

RRID

AB_3071839

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:50

  • IF-Tissue

  • 1:1,000

Target

Function

HLA class II histocompatibility antigen gamma chain also known as HLA-DR antigens-associated invariant chain or CD74 (Cluster of Differentiation 74), is a protein that in humans is encoded by the CD74 gene. The invariant chain (Abbreviated Ii) is a polypeptide which plays a critical role in antigen presentation. It is involved in the formation and transport of MHC class II peptide complexes for the generation of CD4+ T cell responses. The cell surface form of the invariant chain is known as CD74. CD74 is a cell surface receptor for the cytokine macrophage migration inhibitory factor (MIF). The nascent MHC class II protein in the rough endoplasmic reticulum (RER) binds a segment of the invariant chain (Ii; a trimer) in order to shape the peptide-binding groove and prevent the formation of a closed conformation. The invariant chain also facilitates the export of MHC class II from the RER in a vesicle. The signal for endosomal targeting resides in the cytoplasmic tail of the invariant chain. This fuses with a late endosome containing the endocytosed antigen proteins (from the exogenous pathway). Binding to Ii ensures that no antigen peptides from the endogenous pathway meant for MHC class I molecules accidentally bind to the groove of MHC class II molecules. The Ii is then cleaved by cathepsin S (cathepsin L in cortical thymic epithelial cells), leaving only a small fragment called CLIP remaining bound to the groove of MHC class II molecules. The rest of the Ii is degraded. CLIP blocks peptide-binding until HLA-DM interacts with MHC II, releasing CLIP and allowing other peptides to bind. In some cases, CLIP dissociates without any further molecular interactions, but in other cases the binding to the MHC is more stable. The stable MHC class II + antigen complex is then presented on the cell surface. Without CLIP, MHC class II aggregates disassemble and/or denature in the endosomes, and proper antigen presentation is impaired.

Background References

1. Xu S et al. Anti-CD74 antibodies in spondyloarthritis: A systematic review and meta-analysis. Semin Arthritis Rheum. 2021 Feb

2. Xu S et al. CD74 Correlated With Malignancies and Immune Microenvironment in Gliomas. Front Mol Biosci. 2021 Sep

Subcellular Location

Endoplasmic reticulum membrane, trans-Golgi network, cell membrane, lysosome, endosome.

UNIPROT

SwissProt: P04233 Human

Synonyms

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

Expand

Images

  • Western blot analysis of CD74 on different lysates with Mouse anti-CD74 antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/1,000 dilution.<br /><br />Lane 1: Daudi cell lysate<br />Lane 2: Raji cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 33 kDa<br />Observed band size: 35 kDa<br /><br />Exposure time: 2 minutes;<br /><br />12% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:150,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of CD74 on different lysates with Mouse anti-CD74 antibody (HA601117) at 1/1,000 dilution.

    Lane 1: Daudi cell lysate
    Lane 2: Raji cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 33 kDa
    Observed band size: 35 kDa

    Exposure time: 2 minutes;

    12% 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 (HA601117) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of LOVO cells labeling CD74 with Mouse anti-CD74 antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-CD74 antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/50 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) 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.

    Immunocytochemistry analysis of LOVO cells labeling CD74 with Mouse anti-CD74 antibody (HA601117) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-CD74 antibody (HA601117) at 1/50 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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.

  • Immunohistochemical analysis of paraffin-embedded human B-cell lymphoma tissue with Mouse anti-CD74 antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/5,000 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.

    Immunohistochemical analysis of paraffin-embedded human B-cell lymphoma tissue with Mouse anti-CD74 antibody (HA601117) at 1/5,000 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 (HA601117) at 1/5,000 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.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-CD74 antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/5,000 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.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-CD74 antibody (HA601117) at 1/5,000 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 (HA601117) at 1/5,000 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.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue with Mouse anti-CD74 antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/5,000 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.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue with Mouse anti-CD74 antibody (HA601117) at 1/5,000 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 (HA601117) at 1/5,000 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.

  • Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue with Mouse anti-CD74 antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/5,000 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.

    Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue with Mouse anti-CD74 antibody (HA601117) at 1/5,000 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 (HA601117) at 1/5,000 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.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Immunohistochemical analysis of paraffin-embedded human liver tissue (negative) with Mouse anti-CD74 antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601117" style="font-weight: bold;text-decoration: underline;">HA601117</a>) at 1/5,000 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.

    ☑ Relative expression (RE)

    Immunohistochemical analysis of paraffin-embedded human liver tissue (negative) with Mouse anti-CD74 antibody (HA601117) at 1/5,000 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 (HA601117) at 1/5,000 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.

  • Application: IF-Tissue<br /><br />Species: Human<br /><br />Site: B-cell lymphoma<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/1,000

    Application: IF-Tissue

    Species: Human

    Site: B-cell lymphoma

    Sample: Paraffin-embedded section

    Antibody concentration: 1/1,000

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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metafield image

CD74 Recombinant Rabbit Monoclonal Antibody [JF40-10]

Application: WB,IF-Cell,IF-Tissue,IHC-P,FC

Reactivity: Human

Conjugate: unconjugated