CD25 antibody | P4A10

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Mouse anti Dog CD25:FITC

Mouse anti Dog CD25

Mouse anti Dog CD25:RPE

Product Type
Monoclonal Antibody
Clone
P4A10
Isotype
IgG1
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
MCA5916F F 0.1 mg
MCA5916GA F WB* 0.1 mg
MCA5916PE F 100 Tests
Mouse anti Dog CD25, clone P4A10 recognizes 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 hemopoietic 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.

Product Details

Target Species
Dog
Product Form
Purified IgG conjugated to Fluorescein Isothiocyanate Isomer 1 (FITC) - liquid
Product Form
Purified IgG - liquid
Product Form
Purified IgG conjugated to R. Phycoerythrin (RPE) - lyophilized
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
Preparation
Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
Preparation
Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
Buffer Solution
Phosphate buffered saline
Buffer Solution
Phosphate buffered saline
Buffer Solution
Phosphate buffered saline
Preservative Stabilisers
0.09% Sodium Azide (NaN3)
1% Bovine Serum Albumin
Preservative Stabilisers
0.09% Sodium Azide
Preservative Stabilisers
0.09% Sodium Azide (NaN3)
1% Bovine Serum Albumin
5% Sucrose
Carrier Free
Yes
Immunogen
Enriched and stimulated Canine T cells.
Approx. Protein Concentrations
IgG concentration 0.1 mg/ml
Approx. Protein Concentrations
IgG concentration 1.0 mg/ml
Fusion Partners
Spleen cells from immunised RBF/DnJ mice were fused with cells of the mouse P3-653 myeloma cell line.

Storage Information

Storage
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.
Storage
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.
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.
Shelf Life
18 months from date of despatch
Shelf Life
18 months from date of despatch
Shelf Life
12 months from date of reconstitution

More Information

UniProt
O62802 Related reagents
Entrez Gene
IL2RA Related reagents
GO Terms
GO:0016021 integral to membrane
Regulatory
For research purposes only

Applications of CD25 antibody

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 Name Verified Min Dilution Max Dilution
Flow Cytometry Neat 1/10
ELISA
Flow Cytometry 1/50 1/100
Immunohistology - Frozen
Immunohistology - Paraffin
Immunohistology - Resin
Immunoprecipitation
Western Blotting 1
Flow Cytometry Neat
  1. 1Clone P4A10 produces a double band at approximately 55kDa under non-reducing conditions. A non-specific band may be observed at 116kDa
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.
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.
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.
Flow Cytometry
Use 10ul of the suggested working dilution to label 106 cells in 100ul
Flow Cytometry
Use 10ul of the suggested working dilution to label 1x106 cells in 100ul.
Flow Cytometry
Use 10ul of the suggested working dilution to label 106 cells in 100ul
Western Blotting
Clone P4A10 detects a double band of approximately 55kDa in PMA-IO activated canine PBMC lysates.

Secondary Antibodies Available

Description Product Code Pack Size Applications List Price Quantity
Human anti Mouse IgG1:HRP HCA036P 0.1 mg E
Goat anti Mouse IgG (H/L):Alk. Phos. (Multi Species Adsorbed) STAR117A 0.5 mg E WB
Goat anti Mouse IgG (H/L):DyLight®488 (Multi Species Adsorbed) STAR117D488GA 0.1 mg F IF
Goat anti Mouse IgG (H/L):DyLight®549 (Multi Species Adsorbed) STAR117D549GA 0.1 mg F IF WB
Goat anti Mouse IgG (H/L):DyLight®649 (Multi Species Adsorbed) STAR117D649GA 0.1 mg F IF
Goat anti Mouse IgG (H/L):DyLight®680 (Multi Species Adsorbed) STAR117D680GA 0.1 mg F WB
Goat anti Mouse IgG (H/L):DyLight®800 (Multi Species Adsorbed) STAR117D800GA 0.1 mg F IF WB
Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed) STAR117F 0.5 mg F
Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed) STAR117P 0.5 mg E WB
Goat anti Mouse IgG (Fc):FITC STAR120F 1 mg C F
Goat anti Mouse IgG (Fc):HRP STAR120P 1 mg E WB
Rabbit F(ab')2 anti Mouse IgG:RPE STAR12A 1 ml F
Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed) STAR13B 1 mg C E P RE WB
Goat anti Mouse IgG:FITC (Rat Adsorbed) STAR70 0.5 mg F
Goat anti Mouse IgG:RPE (Rat Adsorbed) STAR76 1 ml F
Goat anti Mouse IgG:HRP (Rat Adsorbed) STAR77 0.5 mg C E P
Goat anti Mouse IgG/A/M:Alk. Phos. STAR87A 1 mg C E WB
Goat anti Mouse IgG/A/M:HRP (Human Adsorbed) STAR87P 1 mg E
Rabbit F(ab')2 anti Mouse IgG:Dylight®800 STAR8D800GA 0.1 mg F IF WB
Rabbit F(ab')2 anti Mouse IgG:FITC STAR9B 1 mg F

Negative Isotype Controls Available

Description Product Code Pack Size Applications List Price Quantity
Mouse IgG1 Negative Control:FITC MCA928F 100 Tests F
Mouse IgG1 Negative Control MCA928 100 Tests F
Mouse IgG1 Negative Control:RPE MCA928PE 100 Tests F

Useful Reagents Available

Description Product Code Pack Size Applications List Price Quantity
Rat anti Dog CD4 MCA1038GA 0.1 mg C F
Rat anti Dog CD8 MCA1039GA 0.1 mg C F IF
Mouse anti Dog CD3:FITC MCA1774F 0.1 mg F
Mouse anti Dog CD3 MCA1774GA 0.1 mg C F IF IP P*
Rat anti Dog CD4 MCA1038GA 0.1 mg C F
Rat anti Dog CD8 MCA1039GA 0.1 mg C F IF
Mouse anti Dog CD3:FITC MCA1774F 0.1 mg F
Mouse anti Dog CD3 MCA1774GA 0.1 mg C F IF IP P*
Rat anti Dog CD4 MCA1038GA 0.1 mg C F
Rat anti Dog CD8 MCA1039GA 0.1 mg C F IF
Mouse anti Dog CD3:FITC MCA1774F 0.1 mg F
Mouse anti Dog CD3 MCA1774GA 0.1 mg C F IF IP P*

Application Based External Images

Flow Cytometry

Western Blotting

Product Specific References

References for CD25 antibody

  1. Abrams, V.K. et al. (2010) A novel monoclonal antibody specific for canine CD25 (P4A10): selection and evaluation of canine Tregs.
    Vet Immunol Immunopathol. 135 (3-4): 257-65.
  2. Finn, J.D. et al. (2010) Eradication of neutralizing antibodies to factor VIII in canine hemophilia A after liver gene therapy.
    Blood.116: 5842-8.

Further Reading

  1. Rissetto, K.C. et al. (2010) Cloning and expression of canine CD25 for validation of an anti-human CD25 antibody to compare T regulatory lymphocytes in healthy dogs and dogs with osteosarcoma.
    Vet Immunol Immunopathol. 135 (1-2): 137-45.
  2. Pinheiro, D. et al. (2011) Phenotypic and functional characterization of a CD4(+) CD25(high) FOXP3(high) regulatory T-cell population in the dog.
    Immunology. 132 (1): 111-22.
  3. Bauer, T.R. Jr et al. (2009) Potential large animal models for gene therapy of human genetic diseases of immune and blood cell systems.
    ILAR J. 50 (2): 168-86.
  4. Paoloni, M. & Khanna, C. (2008) Translation of new cancer treatments from pet dogs to humans.
    Nat Rev Cancer. 8 (2): 147-56.