PRODUCT DETAILS

Host: Mouse

Isotype: IgG2a, kappa

Clonality: Monoclonal

Clone: CHI2S4N

Format: PerCP-eFluor 710

Reactivity: Hu, Ms

Application: Flow Cytometry

Tested Dilution: 5 µL (0.03 µg)/test

Concentration: 5 μL/Test

Storage: 4°C, store in dark, DO NOT FREEZE!

Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2

Purification: Affinity chromatography

Data Sheet: TDS

Specific Information

Description: This CHI2S4N monoclonal antibody recognizes human and mouse signal transducer and activator of transcription 6 (STAT6) when phosphorylated on tyrosine 641. Following their phosphorylation by JAKs, STAT proteins translocate to the nucleus where they bind to DNA and regulate transcription of specific genes in a cell type- and cytokine-specific manner. In response to IL-4, STAT6 is phosphorylated on tyrosine 641 by JAK1 and JAK3. STAT6 signaling downstream of the IL-4 receptor promotes T cell growth and B cell production of IgE.

Specificity of this CHI2S4N clone was determined by ELISA and flow cytometry.

Applications Reported: This CHI2S4N antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This CHI2S4N antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 µL (0.03 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.

Staining Protocol: We recommend using Protocol C: Two-step protocol: Fixation/Methanol. Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins and Protocol B: One-step protocol: intracellular (nuclear) proteins cannot be used. All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the BestProtocols® Section under the Resources tab online.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

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: 488 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

 

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