Overview

  • Product name

    Anti-CD45RA antibody [MRC OX-33]
    See all CD45RA primary antibodies
  • Description

    Mouse monoclonal [MRC OX-33] to CD45RA
  • Host species

    Mouse
  • Specificity

    This antibody is directed against a high molecular weight band of the leucocyte common antigen (LCA).

  • Tested applications

    Suitable for: IHC-Fr, Flow Cytmore details
  • Species reactivity

    Reacts with: Rat
  • Immunogen

    Full length native protein (purified) corresponding to Rat CD45RA. Purified spleen leucocyte common antigen.

  • Positive control

    • Flow Cyt: Lewis rat splenocytes. IHC-Fr: Rat Spleen
  • General notes

     

    Spleen cells from immunised BALB/c mice were fused with cells of the NSO/U myeloma cell line. This clone has been described reacting with paraffin embedded material following PLP fixation (see Whiteland et al., 1995). It only labels B cells among thoracic duct lymphocytes, with little labelling in bone marrow and none on thymocytes (Barclay et al., 1987).

     

    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 ab33933 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 µg/ml.
Flow Cyt Use a concentration of 0.2 µg/ml.

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

 

Target

  • Function

    Protein tyrosine-protein phosphatase required for T-cell activation through the antigen receptor. Acts as a positive regulator of T-cell coactivation upon binding to DPP4. The first PTPase domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. Upon T-cell activation, recruits and dephosphorylates SKAP1 and FYN. Dephosphorylates LYN, and thereby modulates LYN activity.
  • Involvement in disease

    Severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive
    Multiple sclerosis
  • Sequence similarities

    Belongs to the protein-tyrosine phosphatase family. Receptor class 1/6 subfamily.
    Contains 2 fibronectin type-III domains.
    Contains 2 tyrosine-protein phosphatase domains.
  • Domain

    The first PTPase domain interacts with SKAP1.
  • Post-translational
    modifications

    Heavily N- and O-glycosylated.
  • Cellular localization

    Membrane. Membrane raft. Colocalized with DPP4 in membrane rafts.
  • Information by UniProt
  • Database links

  • Alternative names

    • B220 antibody
    • CD45 antibody
    • CD45 antigen 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
    • Protein tyrosine phosphatase, receptor type, C antibody
    • Protein tyrosine phosphatase, receptor type, c polypeptide antibody
    • Ptprc antibody
    • PTPRC_HUMAN antibody
    • Receptor-type tyrosine-protein phosphatase C antibody
    • RT7 antibody
    • T200 antibody
    • T200 glycoprotein antibody
    • T200 leukocyte common antigen antibody
    see all

Images

  • Lewis rat splenocytes stained with ab33933 (right) or mouse IgG1κ (ab170190) isotype (left). Lewis rat splenocytes were incubated for 30 min on ice in 1x PBS / 10 % rat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab33933) or mouse IgG1κ isotype (ab170190) (1x106 in 100µl at 0.2 µg/ml) for 30 min on ice.

    The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor ® 488, pre-adsorbed) (ab150177) was used at 1/2000 dilution for 30 min at 4°C. The cells were simultaneously stained with CD3 antibody.

    Acquisition of >30,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on viable lymphocytes.

     

  • IHC image of CD45RA 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 then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M glycine and 1% (w/v) BSA for 1h at room temperature.  The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab33933 at 1µg/ml. The section was then incubated with ab150117 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preabsorbed, (Shown in green) 1/1000) for 1 hour at room temperature. DAPI was used to stain the cell nuclei (blue). The secondary-only control insert image is taken from an identical assay without primary antibody. The section was then mounted using Fluoromount®.

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

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

References

This product has been referenced in:

  • Whiteland JL  et al. Immunohistochemical detection of T-cell subsets and other leukocytes in paraffin-embedded rat and mouse tissues with monoclonal antibodies. J Histochem Cytochem 43:313-20 (1995). Read more (PubMed: 7868861) »
  • Woollett GR  et al. Molecular and antigenic heterogeneity of the rat leukocyte-common antigen from thymocytes and T and B lymphocytes. Eur J Immunol 15:168-73 (1985). Read more (PubMed: 2578966) »
See all 2 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Question
Answer

Thank you for contacting us.

I have emailed the lab to see if they have an IHC-P image they can share, and will let you know when I hear back from them.

The reference I had mentioned is as follows:

Whiteland JL et al. Immunohistochemical detection of T-cell subsets and other leukocytes in paraffin-embedded rat and mouse tissues with monoclonal antibodies. J Histochem Cytochem 43:313-20 (1995). PubMed: 7868861

http://www.ncbi.nlm.nih.gov/pubmed/7868861?dopt=Abstract
Unfortunately, I do not havethe entire paper, only the abstract- it is also not available online. You might need to check in a science library.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
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Question
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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