Anti-CD74 antibody (ab64772)

Overview

  • Product nameAnti-CD74 antibody
    See all CD74 primary antibodies
  • Description
    Rabbit polyclonal to CD74
  • Tested applicationsSuitable for: WB, ICC/IF, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Non Human Primates
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 250 to the C-terminus of Human CD74.

    (Peptide available as ab74386.)

  • Positive control
    • This antibody gave a positive signal in Raji Whole Cell Lysate IF/ICC: Raw246.7 cell line

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferPreservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Associated products

Applications

Our Abpromise guarantee covers the use of ab64772 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IP Use a concentration of 5 µg/ml.

IP image from Phase V. Lot JLD 04.06.2013

IHC-P 1/80 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • FunctionPlays a critical role in MHC class II antigen processing by stabilizing peptide-free class II alpha/beta heterodimers in a complex soon after their synthesis and directing transport of the complex from the endoplasmic reticulum to the endosomal/lysosomal system where the antigen processing and binding of antigenic peptides to MHC class II takes place. Serves as cell surface receptor for the cytokine MIF.
  • Sequence similaritiesContains 1 thyroglobulin type-1 domain.
  • Cellular localizationCell membrane. Endoplasmic reticulum membrane. Golgi apparatus > trans-Golgi network. Endosome. Lysosome. Transits through a number of intracellular compartments in the endocytic pathway. It can either undergo proteolysis or reach the cell membrane.
  • Information by UniProt
  • Database links
  • 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
    see all

Anti-CD74 antibody images

  • CD74 was immunoprecipitated using 0.5mg Raji whole cell extract, 5µg of Rabbit polyclonal to CD74 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Raji whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab64772.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 34kDa; CD74

  • Anti-CD74 antibody (ab64772) at 1 µg/ml + Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

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


    Exposure time : 2 minutes
  • ab64772 (1:160) staining CD74 in paraffin-embedded human tonsil (left panel) using an automated system (Ventana Discovery). Right-hand panel shows negative control (no primary antibody).
    Using this protocol there is strong membrane staining of activated B cells in the germinal centres and B cells of the mantle zone of the follicles plus scattered cells of the interfollicular areas (paracortical B cells).

    Sections were rehydrated and antigen retrieved in CC1 Cell Conditioning Buffer using Ventana Mild Retrieval programme. Slides were blocked in 3% H2O2 / 4 min / 37°C and incubated with ab64772 (1:160 dilution / 2 hours / 37°C). Sections then blocked (4mins / 37°C) and incubated with Dako swine anti-rabbit antibody (1:50, 28 min / 37°C). Staining was amplified and detected by incubation with Ventana Streptavidin ABC system (16 min / 37°C) and Ventana DAB map reagent (8 min / 37°C). Slides were counterstained with Haematox

  • ab64772 (1:80) staining CD74 in paraffin-embedded human lymph node (left panel) using an automated system (Ventana Discovery). Right-hand panel shows negative control (no primary antibody).
    Using this protocol there is strong membrane staining of activated B cells in the germinal centres and B cells of the mantle zone of the follicles plus scattered cells of the interfollicular areas (paracortical B cells).

    Sections were rehydrated and antigen retrieved in CC1 Cell Conditioning Buffer using Ventana Mild Retrieval programme. Slides were blocked in 3% H2O2 / 4 min / 37°C and incubated with ab64772 (1:80 dilution / 1 hour / 37°C). Sections then blocked (3mins / 37°C) and incubated with Dako swine anti-rabbit antibody (1:50, 28 min / 37°C). Staining was amplified and detected by incubation with Ventana Streptavidin ABC system (16 min / 37°C) and Ventana DAB map reagent (8 min / 37°C). Slides were counterstained with Haematoxyli

  • ab64772 (1/250) staining CD74 in paraffin-embedded Human tonsil tissue.  Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval.  The secondary antibody was goat anti rabbit conjugated to HRP.  For further experimental details please refer to abreview.

    See Abreview

  • ICC/IF image of ab64772 stained RAW246.7 cells. The cells were 4% paraformaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab64772, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-CD74 antibody (ab64772)

ab64772 has not yet been referenced specifically in any publications.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Tonsil)
Specification Tonsil
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Dako Target Retrieval solution
Permeabilization No
Blocking step 5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
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Verified customer

Submitted Apr 13 2010

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