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CD25 antibody | K231.3B2

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Anti Pig CD25 Antibody, clone K231.3B2 thumbnail image 1 Anti Pig CD25 Antibody, clone K231.3B2 thumbnail image 2 Anti Pig CD25 Antibody, clone K231.3B2 thumbnail image 3 Anti Pig CD25 Antibody, clone K231.3B2 thumbnail image 4
Anti Pig CD25 Antibody, clone K231.3B2 gallery image 1
Porcine peripheral blood lymphocytes stained with Mouse anti Pig CD25 antibody, clone K231.3B2 (MCA1736GA) followed by FITC conjugated Goat anti Mouse IgG antibody (STAR117F), dual stained with RPE conjugated Mouse anti Pig CD4a antibody, clone MIL17 (MCA1749PE). Numbers indicate the percent cells in each quadrant.
Anti Pig CD25 Antibody, clone K231.3B2 gallery image 2
Published customer image:
Mouse anti Pig CD25, clone K231.3B2 (MCA1736) used for the evaluation of CD25 expression on lymphocytes in culture by flow cytometry.
Image caption:
Induction of Tregs from lymphocytes co-cultured with PRRSV-infected MoDCs. (A) Representative flow cytometry profile of lymphocytes following 3 days co-culture of PBMCs with PRRSV-infected MoDCs (upper layer), and only culture with PRRSV-infected PBMCs (lower layer). (B) Percentage of Foxp3+ cells in the gated CD4+CD25+ subpopulations of PBMCs co-cultured with PRRSV-infected MoDC. (C) Percentage of Foxp3+ cells in the gated CD4+ CD25+ subpopulations of PRRSV-infected PBMCs alone. (D) Sorted CD4+CD25high cells. (E) Percentage suppression at the indicated Tregs: peripheral lymphocyte ratios. The lymphocytes exposed to Marc-145 lysate-treated MoDCs were used as negative control. The percentage of suppression was calculated as follows: % suppression = 100 × [1 − (% proliferation w/PRRSV/ % proliferation w/mock)] [35]. Data from three independent experiments. All data analysis was done using one-way ANOVA and significant differences are shown (*P<0.05 and **P< 0.01).

From: Fan B, Liu X, Bai J, Li Y, Zhang Q, Jiang P (2015) The 15N and 46R Residues of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein Enhance Regulatory T Lymphocytes Proliferation.
PLoS ONE 10(9): e0138772.
Under a Creative Commons license
Anti Pig CD25 Antibody, clone K231.3B2 gallery image 3
Figure A. FITC conjugated mouse anti porcine CD4a (MCA1749F) and purified mouse IgG1 isotype control (MCA928) detected with goat anti mouse IgG1PE (STAR132PE). Figure B. FITC conjugated mouse anti porcine CD4a (MCA1749F) and purified mouse anti porcine CD25 (MCA1736GA) detected with goat anti mouse IgG1PE (STAR132PE). All experiments performed on red cell lysed porcine blood gated on mononuclear cells.
Anti Pig CD25 Antibody, clone K231.3B2 gallery image 4
Figure A. Purified mouse anti porcine CD25 (MCA1736GA) detected with goat anti mouse IgG1 PE (STAR132PE) and mouse IgG2b FITC isotype control (MCA691F). Figure B. Purified mouse anti porcine CD25 (MCA1736GA) detected with goat anti mouse IgG1 PE (STAR132PE) and mouse anti porcine CD4a FITC (MCA1749F). All experiments performed on red cell lysed porcine blood gated on mononuclear cells.

Mouse anti Pig CD25:Alexa Fluor® 647

Mouse anti Pig CD25

Product Type
Monoclonal Antibody
Clone
K231.3B2
Isotype
IgG1
Product Code Applications Pack Size List Price Quantity
MCA1736A647
Datasheet
F
SDS
100 Tests/1ml loader

MCA1736GA
Datasheet
C F
SDS
0.1 mg loader

Mouse anti Pig CD25, clone K231.3B2 recognizes porcine CD25, the alpha chain of the interleukin 2 receptor (IL-2Rα), also known as the low affinity Interleukin 2 receptor. The IL-2 receptor exists in three forms, the high affinity heterodimer, the intermediate affinity β monomer and the low affinity α monomer configurations. Clone K231.3B2 was clustered as CD25 at the First International Workshop to Define Swine Cluster of Differentiation (CD) Antigens (Lunney et al. 1994).

Mouse anti pig CD25, clone K231.3B2 immunoprecipitates a protein of ~65-70 kDa from activated lymphocyte preparations (Bailey et al. 1992).

CD25 is a 270 amino acid single pass type I transmembrane glycoprotein containing 2 Sushi domains. Low expression of CD25 is seen on resting peripheral blood mononuclear cells, rapidly up-regulated following stimulation by concanavalin A and phorbol myristate acetate, indicative of its role as an activation antigen (Bullido et al. 1999).

Product Details

Target Species
Pig
Product Form
Purified IgG conjugated to Alexa Fluor 647 - liquid
Product Form
Purified IgG - liquid
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
Preservative Stabilisers
0.09% Sodium Azide (NaN3)
1% Bovine Serum Albumin
Preservative Stabilisers
0.09% Sodium Azide (NaN3)
Carrier Free
Yes
Immunogen
Con A activated porcine peripheral blood lymphocytes.
Approx. Protein Concentrations
IgG concentration 0.05 mg/ml
Approx. Protein Concentrations
IgG concentration 1.0 mg/ml
Fusion Partners
Spleen cells from immunised mice were fused with cells of the mouse P3-X63-Ag.8.653 myeloma cell line.

Storage Information

Storage
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.

Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Storage
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.

Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Guarantee
12 months from date of despatch
Guarantee
12 months from date of despatch

More Information

UniProt
O02733
Entrez Gene
IL2RA
GO Terms
GO:0016021 integral to membrane
GO:0004872 receptor activity
Acknowledgements
This product is provided under an intellectual property licence from Life Technologies Corporation. The transfer of this product is contingent on the buyer using the purchased product solely in research, excluding contract research or any fee for service research, and the buyer must not sell or otherwise transfer this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; (c) manufacturing or quality assurance or quality control, or (d) resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad CA 92008 USA or outlicensing@thermofisher.com
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
Flow Cytometry 1/25 1/200
Immunohistology - Frozen
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 1x106 cells in 100ul
Flow Cytometry
Use 10ul of the suggested working dilution to label 1x106 cells in 100ul
Copyright © 2021 Bio-Rad Antibodies (formerly AbD Serotec)

Secondary Antibodies Available

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

Negative Isotype Controls Available

Description Product Code Applications Pack Size List Price Quantity
Mouse IgG1 Negative Control:Alexa Fluor® 647 MCA928A647 F 100 Tests/1ml loader
Mouse IgG1 Negative Control MCA928 F 100 Tests loader

Useful Reagents Available

Description Product Code Applications Pack Size List Price Quantity
Mouse anti Pig CD14:FITC MCA1218F F 0.1 mg loader
Mouse anti Pig CD45:Alexa Fluor® 647 MCA1222A647 F 100 Tests/1ml loader
Mouse anti Pig wCD8 Alpha:FITC MCA1223F F 0.1 mg loader
Mouse anti Pig wCD8 Alpha:RPE MCA1223PE F 100 Tests loader
Mouse anti Pig CD4 Alpha:FITC MCA1749F F 0.1 mg loader
Mouse anti Pig CD4 Alpha:RPE MCA1749PE F 100 Tests loader
Mouse anti Pig CD14:FITC MCA1218F F 0.1 mg loader
Mouse anti Pig CD45:Alexa Fluor® 647 MCA1222A647 F 100 Tests/1ml loader
Mouse anti Pig wCD8 Alpha:FITC MCA1223F F 0.1 mg loader
Mouse anti Pig wCD8 Alpha:RPE MCA1223PE F 100 Tests loader
Mouse anti Pig CD4 Alpha:FITC MCA1749F F 0.1 mg loader
Mouse anti Pig CD4 Alpha:RPE MCA1749PE F 100 Tests loader

Application Based External Images

Immunofluorescence

Product Specific References

References for CD25 antibody

  1. Bailey, M. et al. (1992) A monoclonal antibody recognising an epitope associated with pig interleukin-2 receptors.
    J Immunol Methods. 153 (1-2): 85-91.
  2. Silva-Campa, E. et al. (2010) European genotype of porcine reproductive and respiratory syndrome (PRRSV) infects monocyte-derived dendritic cells but does not induce Treg cells.
    Virology. 396 (2): 264-71.
  3. Kick, A.R. et al. (2011) Evaluation of peripheral lymphocytes after weaning and vaccination for Mycoplasma hyopneumoniae.
    Res Vet Sci. 91 (3): e68-72.
  4. Silva-Campa, E. et al. (2009) Induction of T helper 3 regulatory cells by dendritic cells infected with porcine reproductive and respiratory syndrome virus.
    Virology. 387: 373-9.
  5. Leroith, T. et al. (2011) A modified live PRRSV vaccine and the pathogenic parent strain induce regulatory T cells in pigs naturally infected with Mycoplasma hyopneumoniae.
    Vet Immunol Immunopathol.140: 312-6.
  6. Barker, E. et al. (2006) The larynx as an immunological organ: immunological architecture in the pig as a large animal model.
    Clin Exp Immunol. 143: 6-14.
  7. Kuo, Y.R. et al. (2011) Prolongation of composite tissue allotransplant survival by treatment with bone marrow mesenchymal stem cells is correlated with T-cell regulation in a swine hind-limb model.
    Plast Reconstr Surg. 127: 569-79.
  8. Young, D. et al. (2012) Soy-derived di- and tripeptides alleviate colon and ileum inflammation in pigs with dextran sodium sulfate-induced colitis.
    J Nutr. 142: 363-8.
  9. Chattha, K.S. et al. (2013) Divergent immunomodulating effects of probiotics on T cell responses to oral attenuated human rotavirus vaccine and virulent human rotavirus infection in a neonatal gnotobiotic piglet disease model.
    J Immunol. 191: 2446-56.
  10. Fan, B. et al. (2015) The 15N and 46R Residues of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein Enhance Regulatory T Lymphocytes Proliferation.
    PLoS One. 10 (9): e0138772.
  11. Williams, A.R. et al. (2016) Polymerization-dependent activation of porcine γδ T-cells by proanthocyanidins.
    Res Vet Sci. 105: 209-15.
  12. Suradhat, S. et al. (2016) Transdermal delivery of plasmid encoding truncated nucleocapsid protein enhanced PRRSV-specific immune responses.
    Vaccine. 34 (5): 609-15.
  13. Pan, H. et al. (2016) Lymphodepletive effects of rabbit anti-pig thymocyte globulin in neonatal swines.
    Transpl Immunol. 39: 74-83.
  14. Ferrari, L. et al. (2016) Phenotypic modulation of porcine CD14+ monocytes, natural killer/natural killer T cells and CD8αβ+ T cell subsets by an antibody-derived killer peptide (KP).
    Res Vet Sci. 109: 29-39.
  15. Sirisereewan, C. et al. (2017) Positive immunomodulatory effects of heterologous DNA vaccine- modified live vaccine, prime-boost immunization, against the highly-pathogenic PRRSV infection.
    Vet Immunol Immunopathol. 183: 7-15.
  16. Singleton, H. et al. (2016) Establishing Porcine Monocyte-Derived Macrophage and Dendritic Cell Systems for Studying the Interaction with PRRSV-1.
    Front Microbiol. 7: 832.
  17. An, C.H. et al. (2018) Plant synthetic GP4 and GP5 proteins from porcine reproductive and respiratory syndrome virus elicit immune responses in pigs.
    Planta. 247 (4): 973-85.
  18. Nedumpun, T. et al. (2019) Negative Immunomodulatory Effects of Type 2 Porcine Reproductive and Respiratory Syndrome Virus-Induced Interleukin-1 Receptor Antagonist on Porcine Innate and Adaptive Immune Functions.
    Front Immunol. 10: 579.

Further Reading

  1. Piriou-Guzylack, L. (2008) Membrane markers of the immune cells in swine: an update.
    Vet Res. 39: 54.

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