Anti-CD74 antibody [PIN.1] (ab22603)
![Western blot - Anti-CD74 antibody [PIN.1] (ab22603)](https://www.abcam.com/ps/products/22/ab22603/Images/ab22603-450516-anti-cd74-antibody-pin1-western-blot-wildtype-raji-cd74-edited-raji-jurkat-hepg2.jpg "Western blot - 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
- 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
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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. -
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.
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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
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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
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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.
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| ICC/IF |
Use a concentration of 10 µg/ml.
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| Notes |
|---|
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IHC-P
Use at an assay dependent concentration. |
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ICC/IF
Use a concentration of 10 µg/ml. |
Target
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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
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Database links
- Entrez Gene: 972 Human
- Omim: 142790 Human
- SwissProt: P04233 Human
- Unigene: 436568 Human
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Alternative names
- CD 74 antibody
- CD74 antibody
- CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated) antibody
see all
Images
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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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody [PIN.1] (ab22603)](https://www.abcam.com/ps/products/22/ab22603/Images/ab22603-32330-anti-cd74-antibody-pin1-immunohistochemistry.jpg)
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)](https://www.abcam.com/ps/products/22/ab22603/Images/ab22603-197842-anti-cd74-antibody-pin1-immunofluorescence.jpg)
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
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![Immunocytochemistry/ Immunofluorescence - Anti-CD74 antibody [PIN.1] (ab22603)](https://www.abcam.com/ps/products/22/ab22603/Images/ab22603-308966-anti-cd74-antibody-pin1-immunofluorescence.jpg)
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
Datasheets and documents
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Datasheet download
References (10)
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
