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    cd74-antibody-pin1-ab22603.pdf

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Immunology Adaptive Immunity B Cells CD
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Anti-CD74 antibody [PIN.1] (ab22603)

  • Datasheet
Submit a review Q&A (2)References (10)

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Western blot - Anti-CD74 antibody [PIN.1] (ab22603)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [PIN.1] (ab22603)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD74 antibody [PIN.1] (ab22603)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD74 antibody [PIN.1] (ab22603)

Key features and details

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

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-CD74 antibody [EPR4064] (ab108393)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

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/IFmore details
    Unsuitable for: Flow Cyt
  • 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

    • WB: Raji and Human B lymphoblastoid cell lysate. IF/ICC: HepG2 cell line.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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

    Preservative: 0.09% Sodium azide
    Constituents: PBS, 50% Glycerol (glycerin, glycerine)
  • 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

    • Immunology
    • Adaptive Immunity
    • B Cells
    • CD
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • KO cell lines

    • Human CD74 knockout Raji cell line (ab273378)
  • KO cell lysates

    • Human CD74 knockout Raji cell lysate (ab275529)
  • Recombinant Protein

    • Recombinant Human CD74 protein (ab156328)

Applications

The Abpromise guarantee

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.
Notes
IHC-P
Use at an assay dependent concentration.
ICC/IF
Use a concentration of 10 µg/ml.
Application notes
Is unsuitable for Flow Cyt.

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.
  • Target information above from: UniProt accession P04233 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 972 Human
    • Omim: 142790 Human
    • SwissProt: P04233 Human
    • Unigene: 436568 Human
    • 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

    • Western blot - Anti-CD74 antibody [PIN.1] (ab22603)
      Western blot - Anti-CD74 antibody [PIN.1] (ab22603)
      All lanes : Anti-CD74 antibody [PIN.1] (ab22603) at 1 µg/ml

      Lane 1 : Wild-type Raji cell lysate
      Lane 2 : CD74 CRISPR/Cas9 edited Raji cell lysate
      Lane 3 : Jurkat cell lysate
      Lane 4 : HepG2 cell lysate

      Lysates/proteins at 30 µg per lane.

      Performed under reducing conditions.

      Observed band size: 35 kDa why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab22603 observed at 35 kDa. Red - loading control, ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

      ab22603 was shown to react with CD74 in western blot. The band observed in CD74 CRISPR/Cas9 edited cell line ab273378 (CRISPR/Cas9 edited lysate ab275529) below 35 kDa is likely to represent a truncated form. This has not been investigated further. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab22603 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

    • 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. 

    Protocols

    • Flow cytometry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (10)

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

    ab22603 has been referenced in 10 publications.

    • Vainer ED  et al. Novel Proteome Extraction Method Illustrates a Conserved Immunological Signature of MSI-H Colorectal Tumors. Mol Cell Proteomics 19:1619-1631 (2020). PubMed: 32641473
    • Zhou Y  et al. Single-cell RNA landscape of intratumoral heterogeneity and immunosuppressive microenvironment in advanced osteosarcoma. Nat Commun 11:6322 (2020). PubMed: 33303760
    • 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
    • Goyos A  et al. A Distinctive Cytoplasmic Tail Contributes to Low Surface Expression and Intracellular Retention of the Patr-AL MHC Class I Molecule. J Immunol 195:3725-36 (2015). PubMed: 26371256
    • 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

    Show All Reviews Q&A
    Submit a review Submit a question

    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.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 28 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Sep 28 2012

    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.

    Read More

    Abcam community

    Verified customer

    Asked on May 21 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on May 21 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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