Immunofluorescent analysis of CD25 (green) showing staining in the cytoplasm of Jurkat cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CD25 monoclonal antibody (Product # M10AR) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||p55 alpha chain of the human IL-2 receptor|
|Storage buffer||PBS with 0.5% BSA|
|Contains||0.1% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||Assay Dependent|
|Immunohistochemistry (IHC)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Flow Cytometry (Flow)||See 1 publications below|
M10AR targets CD25 in ICC/IF, FACS, IHC, IP, and WB applications and shows reactivity with Human samples.
The M10AR immunogen is p55 alpha chain of the human IL-2 receptor.
M10AR detects CD25 which has a predicted molecular weight of approximately 28 kDa.
The interleukin 2 (IL2) receptor alpha (IL2RA) and beta (IL2RB) chains, together with the common gamma chain (IL2RG), constitute the high-affinity IL2 receptor. Homodimeric alpha chains (IL2RA) result in low-affinity receptor, while homodimeric beta (IL2RB) chains produce a medium-affinity receptor. Normally an integral-membrane protein, soluble IL2RA has been isolated and determined to result from extracellular proteolyisis. Alternately-spliced IL2RA mRNAs have been isolated, but the significance of each is presently unknown. Mutations in this gene are associated with interleukin 2 receptor alpha deficiency.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Heterogeneous immunophenotype of granular lymphocyte expansions: differential expression of the CD8 alpha and CD8 beta chains.
M10AR was used in flow cytometry to investigate the granular lymphocyte expansions with specific monoclonal antibodies
|de Totero D,Tazzari PL,DiSanto JP,di Celle PF,Raspadori D,Conte R,Gobbi M,Ferrara GB,Flomenberg N,Lauria F||Blood (80:1765)||1992|