Description: The H1.2F3 monoclonal antibody reacts with mouse CD69, also known as very early activation antigen (VEA). CD69 is approximately 35 kDa and is expressed on the surface as a disulfide-linked dimer. While a small subset of lymphocytes in the thymus, spleen and lymph nodes express this antigen, activation of both T and B cells rapidly upregulates the surface expression of CD69, suggesting a role for CD69 in lymphocyte development and activation.

Applications Reported: This H1.2F3 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This H1.2F3 antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to 0.5 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Brilliant Ultra Violet™ 496 (BUV496) is a tandem dye that emits at 496 nm and is intended for use on cytometers equipped with an ultraviolet (355 nm) laser. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401-42) or Brilliant Stain Buffer™ (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

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 # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) 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: 350 nm; Emission: 496 nm; Laser: Ultraviolet Laser.

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