Anti-CD45RC antibody [MRC OX-22] (ab33945)

Reviews (1)Q&A (1)References (2)

Overview

  • Product name

    Anti-CD45RC antibody [MRC OX-22]
    See all CD45RC primary antibodies

  • Description

    Mouse monoclonal [MRC OX-22] to CD45RC

  • Host species

    Mouse

  • Specificity

    ab33945 recognises rat CD45RC, the high molecular weight form of the leucocyte common antigen. The antigen is found on B cells, approximately 50% of bone marrow cells, all CD8+ve T cells, but splits CD4+ve T cells into two populations, CD4+CD45RChigh (Th1-like) and CD4+CD45RClow (Th2-like).

  • Tested applications

    Suitable for: IHC-Fr, IHC-P, Flow Cytmore details

  • Species reactivity

    Reacts with: Rat

  • Immunogen

    Tissue, cells or virus corresponding to Rat CD45RC. PHA stimulated rat lymphocytes.

  • Positive control

    • IHC-Fr: Frozen rat spleen. IHC-P: Rat spleen tissue.

  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

Properties

Applications

Our Abpromise guarantee covers the use of ab33945 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-Fr Use a concentration of 1 mg/ml.

This product gave a positive signal in HeLa cells fixed with 10% formaldehyde (10 min).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

Target

  • Relevance

    The CD45 family of cell surface glycoprotein antigens exist as a set of isoforms. The patterns of monoclonal antibody recognition of the CD45 phenotypes can be restricted (CD45RA, RB and RC), or broader, such as the CD45 T 200 species. The "restricted" subsets of this group arise as a result of translation of alternatively spliced mRNAs from exons A, B and C (exons 4, 5 and 6), where exons 3-15 encode the extracellular domain of the CD45 molecule. The restricted epitopes are differentially expressed on T and B lymphocytes, including functionally distinct T cells. CD45 (LCA) is a transmembrane phosphotyrosine phosphatase expressed on leucocytes. Myeloid cells do not express the RC isoform.

  • Cellular localization

    Membrane; Single-pass type I membrane protein. Membrane raft. Note: Colocalized with DPP4 in membrane rafts.

  • Database links

    • Alternative names

      • B220 antibody
      • CD45 antibody
      • CD45R antibody
      • EC 3.1.3.48 antibody
      • GP180 antibody
      • L-CA antibody
      • LCA antibody
      • Leukocyte common antigen antibody
      • loc antibody
      • Ly-5 antibody
      • LY5 antibody
      • Ly5, homolog of antibody
      • Lymphocyte antigen 5 antibody
      • Lyt-4 antibody
      • Protein tyrosine phosphatase receptor type C antibody
      • PTPRC antibody
      • Receptor-type tyrosine-protein phosphatase C antibody
      • RT7 antibody
      • T200 antibody
      • T200 Glycoprotein antibody
      see all

    Images

    • Immunohistochemistry (Frozen sections) - Anti-CD45RC antibody [MRC OX-22] (ab33945)
      Immunohistochemistry (Frozen sections) - Anti-CD45RC antibody [MRC OX-22] (ab33945)

      IHC image of CD45RC staining in a section of frozen normal rat spleen*.

      The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining.

      Non-specific protein-protein interactions were blocked using TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1 hour at room temperature. The section was then incubated with ab33945 (1µg/ml dilution) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. The section was then incubated with ab150119 ((Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647) (shown in red) and DAPI (staining nuclear DNA) (shown in blue) for 1 hour at room temperature. The section was then mounted using Fluoromount®.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

      The DAPI only control (no antibody) inset shows no autofluorescence, demonstrating that any Alexa Fluor® 647 signal is derived directly from bound ab33945.

      For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

      *Tissue obtained from Charles River.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45RC antibody [MRC OX-22] (ab33945)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45RC antibody [MRC OX-22] (ab33945)

      IHC image of CD45RC staining in a section of formalin-fixed paraffin-embedded normal rat spleen performed on a Leica BOND™ system using the standard Protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab33945, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX.

      The inset secondary-only control image is taken from an identical assay without primary antibody.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times

    References

    This product has been referenced in:

    • Arthur RP & Mason D T cells that help B cell responses to soluble antigen are distinguishable from those producing interleukin 2 on mitogenic or allogeneic stimulation. J Exp Med 163:774-86 (1986). Read more (PubMed: 2936864) »
    • Spickett GP  et al. MRC OX-22, a monoclonal antibody that labels a new subset of T lymphocytes and reacts with the high molecular weight form of the leukocyte-common antigen. J Exp Med 158:795-810 (1983). Read more (PubMed: 6224884) »
    See all 2 Publications for this product

    Publishing research using ab33945? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    Question

    Could you provide an IHC-P protocol for ab33933 and ab33934.

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    Answer

    Thank you for your recent telephone enquiry. I have obtained the following general testing method from the originator of ab33933 and ab33934. Please note that this protocol is a guideline only and may need to be optimized in your laboraotry. I hope this will be helpful to you. Should you have any further questions, please do not hesitate to contact us. 1. De-paraffinise sections thoroughly in Xylene/synthetic solvent and hydrate through graded series of alcohols. 2. Wash twice in TBS. 3. If required, treat with 0.3% (w/v) hydrogen peroxide in methanol for 30 minutes to block endogenous peroxidase activity (2% w/v H2O2/Methanol can be used for a shorter time if preferred). 4. Wash once in TBS. 5. If required, include appropriate antigen retrieval step to enhance immunostaining. 6. Wash once in TBS. 7. Incubate sections for 10 minutes in 10% normal serum from species in which secondary antibody was raised. 8. Tap excess serum off the slides before staining. 9. Incubate sections in primary antibody for at least 1 hour at room temperature in a humid chamber or overnight at 4°C. Wash three times in TBS. 10. Add enzyme conjugated secondary antibody at recommended dilution (see specific datasheet for details). Incubate for at least 30 minutes at room temperature. 11. Wash three times in TBS. 12. Incubate in appropriate substrate solution for recommended period of time (we would recommends the use of DAB substrate with HRP conjugated antibodies, and Fast Red / Napthol AS-MX for Alkaline Phosphatase conjugated antibodies). 13. Wash once in water. 14. Counterstain in haematoxylin, 2-10 minutes. ‘Blue’ in running water for 5 minutes. 15. Mount in aqueous mounting medium or alternatively dehydrate through alcohols and xylene/solvent and mount in synthetic mountant. Notes: NB. Note that certain substrates are soluble in alcohol - please refer to supplier information for details. Appropriate control samples should always be included. It may be useful to include a control in which no primary antibody is used at all, to determine any non-specific binding of the secondary reagent to the target tissue. Solutions used: TBS (stock solution ×10 concentrated) Sodium chloride 87.66 g, Tris 60.55 g, Distilled water 1 litre. Adjust pH to 7.5 using concentrated HCl.

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