CD25 Antibody | P4A10

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CD25 Antibody | P4A10 gallery image 1

Canine peripheral blood lymphocytes stained with Mouse anti Dog CD25 (MCA5916GA) followed by Goat anti Mouse IgG:FITC (STAR117F) and dual stained with Rat anti Dog CD4:Alexa Fluor®647 (MCA1038A647)

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CD25 Antibody | P4A10 gallery image 2

Canine peripheral blood lymphocytes stained with Mouse anti Dog CD25:FITC (MCA5916F) and dual stained with Rat anti Dog CD4:Alexa Fluor®647 (MCA1038A647)

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CD25 Antibody | P4A10 gallery image 3

Canine peripheral blood lymphocytes stained with Mouse anti Dog CD25:RPE (MCA5916PE) and dual stained with Rat anti Dog CD4:Alexa Fluor®647 (MCA1038A647)

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CD25 Antibody | P4A10 gallery image 4

Figure A. FITC conjugated Rat anti Canine CD11/18 (MCA1040F) and RPE conjugated Mouse IgG1 isotype control (MCA928PE). Figure B. FITC conjugated Rat anti Canine CD11/18 (MCA1040F) and RPE conjugated Mouse anti Canine CD25 (MCA5916PE). All experiments performed on red cell lysed canine blood gated on mononuclear cells.

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CD25 Antibody | P4A10 gallery image 5

Figure A. PE conjugated Mouse anti Canine CD25 (MCA5916F) and A488 conjugated Rat IgG2c isotype control (MCA2879A488). Figure B. PE conjugated Mouse anti Canine CD25 (MCA5916F) and A488 conjugated Rat anti Canine CD11/18 (MCA1040PE). All experiments performed on red cell lysed canine blood gated on mononuclear cells.

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  • Mouse anti Dog CD25:RPE
  • Mouse anti Dog CD25
  • Mouse anti Dog CD25:FITC
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  • Product Type
    Monoclonal Antibody
  • Clone
    P4A10
  • Isotype
    IgG1
3 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    MCA5916GAF, WB*datasheet pdfdatasheet pdf0.1 mg
    MCA5916GA
    MCA5916FFdatasheet pdfdatasheet pdf0.1 mg
    MCA5916F
    MCA5916PEFdatasheet pdfdatasheet pdf100 Tests
    MCA5916PE
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Mouse anti Dog CD25, clone P4A10 recognises the canine homologue of the human CD25 cell surface antigen, also known at IL-2Rα, a glycoprotein of approximately 55 kDa expressed primarily by activated T lymphocytes (Abrams et al. 2010).

      The IL-2 receptor is composed of 3 subunits, an α chain (CD25), a β chain (CD122) and a γ chain (CD132), CD25 functions as a low affinity receptor for IL-2.

      Research has demonstrated that antibodies to CD4, FoxP3 and CD25 may be used in studies of T regulatory cells (Tregs), a unique subset of T helper cells that function in the control of effector cells vital in preventing autoimmunity.

      Previous studies of canine Tregs have utilized the mouse anti human CD25 monoclonal antibody, clone ACT1, which has demonstrated cross reactivity to canine CD25 (Rissetto et al. 2010). However this clone has poor affinity for canine CD25. The development of clone P4A10 now offers a specific monoclonal Mouse anti Canine CD25, demonstrating a greater affinity for canine CD25 than the cross reactive anti human antibody (Abrams et al. 2010).

      The dog is an important veterinary species in its own right. In addition dogs are used as an animal model in the study of a number of serious human disease states including various forms cancers (Paoloni et al. 2008), and in genetically related diseases of the hemopoitic system (Bauer et al. 2009). The generation of clone P4A10 provides an improved tool for the identification of canine CD25 and will facilitate studies in this species.
    • Intended Use
    • Target Species
      Dog
    • Product Form
      Purified IgG conjugated to R. Phycoerythrin (RPE) - lyophilised
      Purified IgG - liquid
      Purified IgG conjugated to Fluorescein Isothiocyanate Isomer 1 (FITC) - liquid
    • Reconstitution
      Reconstitute with 1.0 ml distilled water
      Care should be taken during reconstitution as the protein may appear as a film at the bottom of the vial. Bio-Rad recommend that the vial is gently mixed after reconstitution.
    • Preparation
      Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
      Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
      Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
    • Preservative Stabilisers
      0.09% Sodium Azide (NaN3)
      1% Bovine Serum Albumin
      5% Sucrose
      0.09% Sodium Azide
      0.09% Sodium Azide (NaN3)
      1% Bovine Serum Albumin
    • Immunogen
      Enriched and stimulated Canine T cells.
    • Purity
    • Approx. Protein Concentrations
      IgG concentration 1.0 mg/ml
      IgG concentration 0.1 mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
      Phosphate buffered saline
      Phosphate buffered saline
    • Fusion Partners
      Spleen cells from immunised RBF/DnJ mice were fused with cells of the mouse P3-653 myeloma cell line.
    • Storage
      Prior to reconstitution store at +4oC.
      After reconstitution store at +4oC.
      DO NOT FREEZE. This product should be stored undiluted. This product is photosensitive and should be protected from light. Should this product contain a precipitate we recommend microcentrifugation before use.
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. This product is photosensitive and should be protected from light. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      12 months from date of reconstitution
      18 months from date of despatch
      18 months from date of despatch
    • GO Terms
      integral to membrane
    • UniProt
    • Entrez Gene
    • Acknowledgements
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      Flow CytometryNeat
    • Application NameYesNoMin DilutionMax Dilution
      ELISA
      Flow Cytometry1/501/100
      Immunohistology - Frozen
      Immunohistology - Paraffin
      Immunohistology - Resin
      Immunoprecipitation
      Western Blotting(1)
      (1)
      Clone P4A10 produces a double band at approximately 55kDa under non-reducing conditions. A non-specific band may be observed at 116kDa
    • Application NameYesNoMin DilutionMax Dilution
      Flow CytometryNeat1/10

    • Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
    • Technical Advice
    • Technical Advice
    • Technical Advice
    • Recommended Protocol
    • Recommended Protocol
    • Recommended Protocol
    • Flow Cytometry
      Use 10ul of the suggested working dilution to label 1x106 cells in 100ul.
    • ELISA
    • ELISA
    • ELISA
    • Immunohistology
    • Immunohistology
    • Immunohistology
    • Histology Positive Control Tissue
    • Histology Positive Control Tissue
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Immunofluorescence
    • Immunofluorescence
    • Western Blotting
    • Western Blotting
      Clone P4A10 detects a double band of approximately 55kDa in PMA-IO activated canine PBMC lysates.
    • Western Blotting
    • Instructions For Use
    • Instructions For Use
    • Instructions For Use

    Additional CD25 Antibody Formats

    Formats Clone Applications Sizes available
    CD25 Antibody : RPE P4A10 F 100 Tests
    CD25 Antibody : FITC P4A10 F 0.1 mg
    CD25 Antibody : Purified P4A10 F, WB* 0.1 mg
    • Copyright © 2016 Bio-Rad Antibodies (formerly AbD Serotec)

    Recommended Secondary Antibody

      DescriptionProduct CodePack SizeApplicationsList PriceQuantity
      Goat anti Mouse IgG (H/L):Alk. Phos. (Multi Species Adsorbed)STAR117A0.5 mgE, WB
      STAR117A
      Goat anti Mouse IgG/A/M:Alk. Phos.STAR87A1 mgC, E, WB
      STAR87A
      Goat anti Mouse IgG (H/L):DyLight®488 (Multi Species Adsorbed)STAR117D488GA0.1 mgF, IF
      STAR117D488GA
      Goat anti Mouse IgG (H/L):DyLight®549 (Multi Species Adsorbed)STAR117D549GA0.1 mgF, IF
      STAR117D549GA
      Goat anti Mouse IgG (H/L):DyLight®649 (Multi Species Adsorbed)STAR117D649GA0.1 mgF, IF
      STAR117D649GA
      Goat anti Mouse IgG (H/L):DyLight®680 (Multi Species Adsorbed)STAR117D680GA0.1 mgF, WB
      STAR117D680GA
      Goat anti Mouse IgG (H/L):DyLight®800 (Multi Species Adsorbed)STAR117D800GA0.1 mgF, IF, WB
      STAR117D800GA
      Rabbit F(ab')2 anti Mouse IgG:Dylight®800STAR8D800GA0.1 mgF, IF, WB
      STAR8D800GA
      Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed)STAR117F0.5 mgF
      STAR117F
      Goat anti Mouse IgG:FITC (Rat Adsorbed)STAR700.5 mgF
      STAR70
      Goat anti Mouse IgG (Fc):FITCSTAR120F1 mgC, F
      STAR120F
      Rabbit F(ab')2 anti Mouse IgG:FITCSTAR9B1 mgF
      STAR9B
      Human anti Mouse IgG1:HRPHCA036P0.1 mgE
      HCA036P
      Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed)STAR117P0.5 mgE, WB
      STAR117P
      Goat anti Mouse IgG:HRP (Rat Adsorbed)STAR770.5 mgC, E, P
      STAR77
      Goat anti Mouse IgG (Fc):HRPSTAR120P1 mgE, WB
      STAR120P
      Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed)STAR13B1 mgC, E, P, RE, WB
      STAR13B
      Goat anti Mouse IgG/A/M:HRP (Human Adsorbed)STAR87P1 mgE
      STAR87P
      Rabbit F(ab')2 anti Mouse IgG:RPESTAR12A1 mlF
      STAR12A
      Goat anti Mouse IgG:RPE (Rat Adsorbed)STAR761 mlF
      STAR76

      Recommended Negative Isotype Control

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse IgG1 Negative Control:RPEMCA928PE100 TestsF
        MCA928PE
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse IgG1 Negative ControlMCA928100 TestsF
        MCA928
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse IgG1 Negative Control:FITCMCA928F100 TestsF
        MCA928F

        Useful Reagents

          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          Mouse anti Dog CD3:FITCMCA1774F0.1 mgF
          MCA1774F
          Rat anti Dog CD4MCA1038GA0.1 mgC, F
          MCA1038GA
          Rat anti Dog CD8MCA1039GA0.1 mgC, F, IF
          MCA1039GA
          Mouse anti Dog CD3MCA1774GA0.1 mgC, F, IF, IP, P *
          MCA1774GA
          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          Mouse anti Dog CD3:FITCMCA1774F0.1 mgF
          MCA1774F
          Rat anti Dog CD4MCA1038GA0.1 mgC, F
          MCA1038GA
          Rat anti Dog CD8MCA1039GA0.1 mgC, F, IF
          MCA1039GA
          Mouse anti Dog CD3MCA1774GA0.1 mgC, F, IF, IP, P *
          MCA1774GA
          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          Mouse anti Dog CD3:FITCMCA1774F0.1 mgF
          MCA1774F
          Rat anti Dog CD4MCA1038GA0.1 mgC, F
          MCA1038GA
          Rat anti Dog CD8MCA1039GA0.1 mgC, F, IF
          MCA1039GA
          Mouse anti Dog CD3MCA1774GA0.1 mgC, F, IF, IP, P *
          MCA1774GA

          Recommended Positive Controls

            Histology Controls

              References

              Further Reading

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