Invitrogen

CD69 Monoclonal Antibody (H1.2F3), eFluor 450, eBioscience™

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CD69 Antibody — Figure 2 Functional screening of thymocyte activation. ( A) Gating strategy and controls. Flow cytometry plots showing active caspase 3 (ActCasp3), CD69, and TCRbeta staining within the thymocyte DP gate. Representative plots for each condition were selected from a set of quadruplicates. NS: Non-stimulated; alpha-CD3/CD28 + DMSO: Samples stimulated with CD3/CD28-coated beads and treated with DMSO; DEX: dexamethasone-treated samples. (B,C) Complete data from the functional screen. Thymocytes were stimulated with anti-CD3/CD28 beads for 18 h, surface stained for CD4, CD8 and CD69, permeabilized, and stained for intracellular active caspase 3. Graphs show the percentage of cells positive for active caspase 3 (ActCasp3; panel B) or CD69 expression (panel C), arranged in order of decreasing value. Raw values (top rows) and values normalized to controls (bottom rows) are shown separately for both active caspase 3 and CD69. The red lines in the raw data charts represent the average value of the DMSO-treated stimulated samples for each experiment. Normalized values are obtained by normalizing individual values to the DMSO-treated stimulated samples. An empirically selected window of +-20% is highlighted in grey in the normalized charts to represent the compounds that were filtered out. Colored symbols represent compounds detailed in Figs 3 and 4 .

CD69 Antibody in Flow Cytometry (Flow) — Staining of unstimulated (left) and onvernight ConA-stimulated (right) C57BL/6 splenocytes with 0.125 µg of Armenian Hamster IgG Isotype Control eFluor® 450 (Product # 48-4888-82) (blue histogram) or 0.125 µg of Anti-Mouse CD69 eFluor® 450 (purple histogram). Total viable cells were used for analysis.

Product Details

Tested Applications

Dilution

Flow Cytometry (Flow)

0.25 µg/test

Published Applications

Flow Cytometry (Flow)

See 23 publications below

Product Specifications

Species Reactivity

Mouse

Published species

Mouse

Host / Isotype

Armenian hamster / IgG

Recommended Isotype Control

Armenian Hamster IgG Isotype Control (eBio299Arm), eFluor 450, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

H1.2F3

Conjugate

eFluor® 450

Excitation/Emission Profile

View spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.1% gelatin

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

RRID

AB_10719430

Product Specific Information

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 24 hour Con A-stimulated mouse splenocytes. This can be used at less than or equal to 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

eFluor® 450 is an alternative to Pacific Blue®. eFluor® 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochome.

Excitation: 405 nm; Emission: 445 nm; Laser: Violet Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

CD69 (AIM, Active Inducer Molecule) is a gp28/34 disulfide bonded homodimer with a molecular weight of 60 kDa under non-reducing conditions. CD69 contains one or two N linked oligosacaride and the molecule is present on activated platelets. In normal peripheral blood a variable percentage of cells express the CD69 antigen, and it is involved in lymphocyte signal transduction. Expression CD69 is induced upon activation of T lymphocytes, and may play a role in proliferation. Furthermore, the protein may act to transmit signals in natural killer cells and platelets. Alternative splicing results in multiple transcript variants of CD69. Diseases associated with CD69 dysfunction include coccidiodomycosis and asthma.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: C-type lectin domain family 2; CD69; CD69 antigen; CLEC2C; Early activation antigen CD69; member C; Very Early Activation Antigen

Gene Aliases: 5830438K24Rik; AI452015; AIM; Cd69; VEA

UniProt ID: (Mouse) P37217

Entrez Gene ID: (Mouse) 12515

Molecular Function: membrane-bound signaling molecule signaling molecule

Process(es)

calcium ion binding carbohydrate binding

CD69 Monoclonal Antibody (H1.2F3), eFluor 450, eBioscience™

Product Details

Product Specific Information

Target Information

Bioinformatics

Material safety data sheets (MSDS)

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