Invitrogen

CD25 Monoclonal Antibody (BC96), Super Bright™ 780, eBioscience™

19 Published Figures

22 References

View all (229) CD25 antibodies

Datasheet

Protocols

Questions & Answers

Datasheet

Protocols

Questions & Answers

Antibody Testing Data (1)
Published Figures (19)

FIGURE: 1 / 20

CD25 Antibody (78-0259-42) in Flow

Staining of normal human peripheral blood cells with CD4 Monoclonal Antibody, APC (Product # 17-0049-42) and Mouse IgG1 kappa Isotype Control, Super Bright 780 (Product # 78-4714-42) (left) or CD25 Monoclonal Antibody, Super Bright 780 (right). Cells in the lymphocyte gate were used for analysis.

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Product Details

78-0259-42

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

5 µL (0.25 µg)/test

View 22 publications 22 publications

Product Specifications

Species Reactivity

Human

Published species

Not Applicable

Host/Isotype

Mouse / IgG1, kappa

Recommended Isotype Control

Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), Super Bright™ 780, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

BC96

Conjugate

Super Bright™ 780 Super Bright™ 780 Super Bright™ 780

Excitation/Emission Max

413/780 nm View spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2724074

Product Specific Information

Description: The BC96 monoclonal antibody reacts with human CD25, the 55 kDa interleukin-2 receptor alpha chain (IL-2R alpha). CD25 is expressed by early progenitors of T and B lineage as well as by activated mature T and B lymphocytes. By itself, CD25 binds IL-2 only with low affinity. However, CD25 associates with CD122 (IL-2 receptor beta chain) and CD132 (common gamma chain) to form the high affinity IL-2 receptor. CD25 plays a role in lymphocyte differentiating and activation/proliferation.

Applications Reported: This BC96 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This BC96 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Super Bright 780 can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter, or equivalent. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.

In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.

Target Information

CD25 (IL2 receptor alpha chain/IL2RA) is a cytokine that plays a role in the proliferation of T and B lymphocytes. The receptor of this cytokine (IL2RA) is a heterotrimeric protein complex with a gamma chain also shared by interleukin 4 (IL4) and interleukin 7 (IL7). IL2RA, IL2R beta chain (IL2RB), and the IL2R gamma chain (IL2RG), constitute the high-affinity IL2 receptor. Homodimeric IL2RA chains result in low-affinity receptor, while homodimeric IL2RB chains produce a medium-affinity receptor. The expression of IL2 in mature thymocytes is monoallelic, which represents an unusual regulatory mode for controlling the precise expression of a single gene. IL2 is primarily produced by mature T cells. IL2 plays an important role as a growth factor, differentiation factor, and regulator of cell death. IL-2 stimulates the proliferation of B cells, augments natural killer cell activity, and inhibits granulocyte macrophage colony formation. The targeted disruption of a similar gene in mice leads to ulcerative colitis-like disease, which suggests a role in the immune response to antigenic stimuli. 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.

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