Intracellular staining of unstimulated (orange histogram) or 15-minute IL-4-stimulated (purple histogram) Th2-polarized CD4+ T cells with Anti-Human/Mouse phospho-STAT6 (Y641) PerCP-eFluor® 710. Cells in the lymphocyte gate were used for analysis.

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Product Details

46-9013-41

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

5 µL (0.03 µg)/test

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Product Specifications

Host/Isotype

Mouse / IgG2a, kappa

Recommended Isotype Control

Mouse IgG2a kappa Isotype Control (eBM2a), PerCP-eFluor™ 710, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

CHI2S4N

Conjugate

PerCP-eFluor™ 710 PerCP-eFluor™ 710 PerCP-eFluor™ 710

Excitation/Emission Max

482/708 nm View spectra

Form

Liquid

Concentration

5 µL/Test

Storage conditions

4°C, store in dark, DO NOT FREEZE!

Shipping conditions

Wet ice

AB_2573861

Product 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.

Target Information

STAT6 protein is a transcription factor activated by cytokines, particularly interleukin-4 and IL13. (Stat6-/-) were found to be deficient in IL-4-mediated functions including Th2 helper T-cell differentiation, expression of cell surface markers, T-cell proliferation, immunoglobulin class switching to IgE, and partial loss of IL-4 mediated proliferation. STAT6 mRNA has been detected in peripheral blood lymphocytes, colon, intestine, ovary, prostate, thymus, spleen, kidney, liver, lung and placenta. STAT6 is critically involved in Th2 immune response. STAT6 is crucial for IL-4-mediated biological responses, and naturally in two isoforms: STAT6a and STAT6c.

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

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Phospho-STAT6 (Tyr641) Monoclonal Antibody (CHI2S4N), PerCP-eFluor™ 710, eBioscience™

Phospho-STAT6 (Tyr641) Antibody (46-9013-41) in Flow