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  1. Link

    cd45rc-antibody-mrc-ox-22-ab33945.pdf

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Immunology Adaptive Immunity B Cells CD
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Anti-CD45RC antibody [MRC OX-22] (ab33945)

  • Datasheet
Reviews (1)Q&A (1)References (3)

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

Key features and details

  • Mouse monoclonal [MRC OX-22] to CD45RC
  • Suitable for: IHC-Fr, IHC-P, Flow Cyt
  • Reacts with: Rat, Human
  • Isotype: IgG1

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488

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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
  • Tested applications

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

    Reacts with: Rat, Human
  • 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.

    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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    MRC OX-22
  • Myeloma

    NS1
  • Isotype

    IgG1
  • Light chain type

    kappa
  • Research areas

    • Immunology
    • Adaptive Immunity
    • B Cells
    • CD
    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Immunology
    • Cell Type Markers
    • CD
    • Non-lineage
    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Phosphatases
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • B Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage

Associated products

  • Alternative Versions

    • Anti-CD45RC antibody [MRC OX-22] - BSA and Azide free (ab244570)
    • Alexa Fluor® 488 Anti-CD45RC antibody [MRC OX-22] (ab256260)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)

Applications

The Abpromise guarantee

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 (1)
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.
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

  • 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.
  • Target information above from: UniProt accession P08575 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5788 Human
    • Entrez Gene: 24699 Rat
    • Omim: 151460 Human
    • SwissProt: P08575 Human
    • SwissProt: P04157 Rat
    • Unigene: 654514 Human
    • Unigene: 90166 Rat
    • 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
      • PTPRC_HUMAN antibody
      • Receptor-type tyrosine-protein phosphatase C antibody
      • RT7 antibody
      • T200 antibody
      • T200 Glycoprotein antibody
      see all

    Images

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

      Flow cytometry staining of Lewis rat splenocytes with ab33945 (right) or mouse IgG1&kappa (ab170190) isotype (left). Cells were incubated for 30 min on ice in 1x PBS containing 10 % rat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab33945) or mouse IgG1&kappa (ab170190) isotype (1x106 in 100 μl; at 0.2 μg/ml) for 30 min on ice.

      The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor ® 488, pre-adsorbed) (ab150117) was used at 1/2000 dilution for 30 min on ice.

      The cells were simultaneously stained with CD3 APC.

      Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on live single cells.

    • 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

    Protocols

    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (3)

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

    ab33945 has been referenced in 3 publications.

    • Yang WI  et al. Blocking the receptor for advanced glycation end product activation attenuates autoimmune myocarditis. Circ J 78:1197-205 (2014). PubMed: 24599045
    • 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). 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). PubMed: 6224884

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a question

    Question

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

    Read More

    Abcam community

    Verified customer

    Asked on Sep 18 2009

    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.

    Read More

    Abcam Scientific Support

    Answered on Sep 18 2009

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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