Spark Blue™ 574 anti-human CD25 (Flexi-Fluor™) Antibody

Product Details

Verified Reactivity

Human

Antibody Type

Monoclonal

Host Species

Mouse

Immunogen

Human CD25 transfected cells

Formulation

Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide

Preparation

The antibody was purified by affinity chromatography and conjugated with Spark Blue™ 574 under optimal conditions.

Concentration

0.2 mg/mL

Storage & Handling

The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.

Application

FC

Recommended Usage

Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.

For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 10⁷ cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.

Excitation Laser

Blue Laser (488 nm)

Application Notes

Clone S20019C can completely block binding of clone BC96 and clone TÜ69 on target cells and can be blocked by BC96 and by TÜ69.

Clone S20019C does not block the binding of clone S20019D and clone M-A251 on target cells.

Clone S20019C can block IL-2 binding to receptor.

Clone S20019C shows comparable CD25 staining on 4% formaldehyde-fixed cells compared to unfixed.

Additional Product Notes

For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.

RRID

AB_3662478 (BioLegend Cat. No. 285138)

Antigen Details

Structure

55 kD transmembrane protein

Distribution

Activated T-Cells, T regulatory Cells, monocytes

Function

IL-2 receptor

Interaction

CD122, CD132

Ligand/Receptor

IL-2

Cell Type

Lymphocytes, T cells, Tregs

Biology Area

Cell Biology, Immunology

Molecular Family

CD Molecules, Cytokine/Chemokine Receptors

Antigen References

1. Schlossman S, et al. Eds. 1995. Leucocyte Typing V. Oxford University Press. New York.
2. Waldmann T. 1991. J Biol. Chem. 266:2681-4.
3. Sakaguchi S, et al. 1995. The Journal of Immunology. 155:1151-64.

Gene ID

NA

UniProt

View information about CD25 on UniProt.org

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol

Other Formats