Anti-CD74 antibody [PIN.1] (ab22603)

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Key features and details

  • Mouse monoclonal [PIN.1] to CD74
  • Suitable for: IHC-P, ICC/IF, WB, IP, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-CD74 antibody [PIN.1]
    See all CD74 primary antibodies

  • Description

    Mouse monoclonal [PIN.1] to CD74

  • Host species

    Mouse

  • Specificity

    This antibody detects an ~33-35 kDa protein doublet, corresponding to the apparent molecular mass of the p33 and p35 forms of human CD74.

  • Tested applications

    Suitable for: IHC-P, ICC/IF, WB, IP, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide:

    DQKPVMDDQRDLISNNE

    conjugated to KLH, corresponding to amino acids 12-28 of Human CD74.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • Epitope

    The epitope is in a region of the cytoplasmic tail of human CD74 which is common to all isoforms.

  • Positive control

    • Human B lymphoblastoid cell lysate. IF/ICC: HepG2 cell line.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped 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 buffer

    pH: 7.20
    Preservative: 0.09% Sodium azide
    Constituents: PBS, 50% Glycerol
  • Concentration information loading...

  • Purity

    Protein G purified

  • Purification notes

    This antibody was purified by immunoaffinity chromatography.

  • Clonality

    Monoclonal

  • Clone number

    PIN.1

  • Isotype

    IgG1

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab22603 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
IHC-P Use at an assay dependent concentration.
ICC/IF Use a concentration of 10 µg/ml.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 33 kDa.
IP Use a concentration of 12 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Plays 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 similarities

    Contains 1 thyroglobulin type-1 domain.

  • Cellular localization

    Cell 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

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [PIN.1] (ab22603)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [PIN.1] (ab22603)
      Ab22603 staining CD74 in human liver. Staining is localized to the membrane and cytoplasm.
      Left panel: with primary antibody at 4ug/ml. Right panel: isotype control.
      Sections were stained using an automated system (Dako Autostainer plus), at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer, EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be req
    • Immunocytochemistry/ Immunofluorescence - Anti-CD74 antibody [PIN.1] (ab22603)
      Immunocytochemistry/ Immunofluorescence - Anti-CD74 antibody [PIN.1] (ab22603)

      ICC/IF image of ab22603 stained HepG2 cells. The cells were 100% methanol fixed (5 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 (ab22603, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse 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

    • Immunocytochemistry/ Immunofluorescence - Anti-CD74 antibody [PIN.1] (ab22603)
      Immunocytochemistry/ Immunofluorescence - Anti-CD74 antibody [PIN.1] (ab22603)

      ICC/IF image of ab22603 staining CD74 in HaCaT cells. Cells were fixed with cold 100% methanol for 10 minutes at -20°C. Samples were incubated with primary antibody at 1:100 dilution for 1 hour at room temperature. A FITC Goat anti-mouse (green) was used as a secondary antibody at 1:50 dilution for 1 hour at room temperature. 

    • Flow Cytometry - Anti-CD74 antibody [PIN.1] (ab22603)
      Flow Cytometry - Anti-CD74 antibody [PIN.1] (ab22603)
      Overlay histogram showing Raji cells stained with ab22603 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab22603, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Raji cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

    References

    Publishing research using ab22603? Please let us know so that we can cite the reference in this datasheet.

    ab22603 has been referenced in 7 publications.

    • Kong XF  et al. Disruption of an antimycobacterial circuit between dendritic and helper T cells in human SPPL2a deficiency. Nat Immunol 19:973-985 (2018). PubMed: 30127434
    • Tato M  et al. Cathepsin S inhibition combines control of systemic and peripheral pathomechanisms of autoimmune tissue injury. Sci Rep 7:2775 (2017). PubMed: 28584258
    • Theron M  et al. Pharmacodynamic Monitoring of RO5459072, a Small Molecule Inhibitor of Cathepsin S. Front Immunol 8:806 (2017). WB ; Human . PubMed: 28769925
    • van de Weijer ML  et al. The E3 Ubiquitin Ligase TMEM129 Is a Tri-Spanning Transmembrane Protein. Viruses 8:N/A (2016). Human . PubMed: 27854284
    • Fernandez-Cuesta L  et al. CD74-NRG1 Fusions in Lung Adenocarcinoma. Cancer Discov 4:415-22 (2014). PubMed: 24469108
    • Landsverk OJ  et al. Invariant chain increases the half-life of MHC II by delaying endosomal maturation. Immunol Cell Biol 89:619-29 (2011). PubMed: 21116285
    • Leth-Larsen R  et al. Metastasis-related plasma membrane proteins of human breast cancer cells identified by comparative quantitative mass spectrometry. Mol Cell Proteomics 8:1436-49 (2009). PubMed: 19321434

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Question

    I am searching for an antibody to CD74 that reacts with human for use in WB and recognizes an N-terminal epitope.

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    Answer

    Thank you for contacting us. I would recommend ab22603, which recognizes a very similar epitope (aa12-28) when compared to the antibody you had been using: ab97479 (aa1-28). I hope this helps, please let me know if you need any additional information or assistance.

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    Question

    : Dear abcam, I need to purchase some anti-CD74 antibody (clone PIN.1 catalog #: ab22603) but I need a custom order. The buffer for this antibody contains 50% glycerol and 0.09% sodium azide. I need to purchase some of this antibody but in a buffer that does NOT contain glycerol nor azide. How quickly can such a custom order be ready? I would kindly need to purchase this as soon as possible.

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    Answer

    Thank you for your reply.

    I spoke to the lab and they have told me that they will be unable to produce a large quantity of ab22603 that is glycerol and azide free. Therefore the best way to get this antibody without any of those reagents present would be to use the antibody concentration kit:

    https://www.abcam.com/Antibody-Concentration-Kit-1-column-ab102776.html

    If there is anything else I can help you with, please let me know.

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