Alexa Fluor® 488 Mouse anti-Mouse Stat6 (pY641)

BD Phosflow™ Alexa Fluor® 488 Mouse anti-Mouse Stat6 (pY641)

Name: Alexa Fluor® 488 Mouse anti-Mouse Stat6 (pY641)
Brand: BD Phosflow™
SKU: 558243

Clone J71-773.58.11

(RUO)

Analysis of Stat6 (pY641) in activated mouse B lymphocytes. Mouse M12 B lymphoma cells were either stimulated with mouse IL-4 (Cat. No. 550067) at 37˚C for 15 minutes (filled histogram) or unstimulated (open histogram). The cells were fixed with BD Phosflow™ Fix Buffer I (Cat. No. 557870) at 37˚C for 10 minutes, permeabilized in BD Phosflow™ Perm Buffer III (Cat. No. 558050) on ice for at least 30 minutes (or overnight at -20˚C), and stained with Alexa Fluor® 488 Mouse anti-Mouse Stat6 (pY641) (Cat. No. 558243). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

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Product Details

Brand: BD Phosflow™

Reactivity: Mouse (QC Testing)

Isotype: Mouse IgG1, κ

Immunogen: Phosphorylated Mouse STAT6 (Y641) Peptide

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Vol. Per Test: 20 µl

RRID: AB_647099

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.

Recommended Assay Procedures

Note: For flow cytometric analysis of human cells, Alexa Fluor® 488 Mouse Anti- Stat6 (pY641) mAb 18 (Cat. No. 612600) is recommended.

Product Notices

This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
An isotype control should be used at the same concentration as the antibody of interest.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Fix Buffer I RUO

Size 250 mL Cat No. 557870

Perm Buffer III RUO

Size 125 mL Cat No. 558050

Alexa Fluor® 488 Mouse IgG1 κ Isotype Control RUO

Size 50 Tests Cat No. 557782

Recombinant Mouse IL-4 RUO

Size 10 µg Cat No. 550067

558243 Rev. 5

Antibody Details

J71-773.58.11

STATs (signal transducers and activators of transcription) are critical mediators of the biologic activity of cytokines including Interleukins (IL) 2-5, IL-7, IL-15, GM-CSF, erythropoietin and growth hormone. Ligand-receptor interaction leads to activation of constitutively associated JAK family kinases and subsequent recruitment/activation of STATs by tyrosine phosphorylation. Active STATs then move to the nucleus to promote transcription of cytokine-inducible genes. Seven STAT proteins have been cloned, each of which is differentially expressed and/or activated in a cytokine-specific and cell type-specific manner. Stat6 plays an important role in signaling pathways that lead to the differentiation of T helper type 2 (Th2) cells from uncommitted CD4 T cell precursors. Moreover, IL-4, secreted by activated T lymphocytes, basophils, and mast cells, induces specific gene expression via the induction of tyrosine phosphorylation of Stat6 at tyrosine 641 (Y641). The SH3:SH2 domain of Stat6 associates with tyrosine-phosphorylated IL-4 receptor and the proximal Jak kinase phosphorylates Stat6 at Y641 on the C-terminal side of the SH2 domain. Stat6 is then released from the receptor, dimerizes, and is thought to contact the basal transcription machinery by binding to p300/CBP. While Stat6 is widely expressed in human tissues, it exhibits elevated expression in peripheral blood lymphocytes, colon, intestine, ovary, prostate, thymus, spleen, kidney, liver, lung, and placenta.

The J71-773.58.11 antibody recognizes mouse Stat6 phosphorylated at Y641.

558243 Rev. 5

Format Details

Alexa Fluor™ 488

Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: Alexa Fluor™ 488

Excitation Source: Blue 488 nm

Excitation Max: 494 nm

Emission Max: 517 nm

558243 Rev.5

Citations & References

Development References (8)

Bromberg J, Darnell JE. The role of STATs in transcriptional control and their impact on cellular function. Oncogene. 2000; 19(21):2468-2473. (Biology). View Reference
Daley JM, Brancato SK, Thomay AA, Reichner JS, Albina JE.. The phenotype of murine wound macrophages.. J Leukoc Biol. 2010; 87(1):59-67. (Clone-specific: Flow cytometry). View Reference
Dent AL, Hu-Li J, Paul WE, Staudt LM. T helper type 2 inflammatory disease in the absence of interleukin 4 and transcription factor STAT6. Proc Natl Acad Sci U S A. 1998; 95(23):13823-13828. (Biology). View Reference
Hale MB, Krutzik PO, Samra SS, Crane JM, Nolan GP.. Stage Dependent Aberrant Regulation of Cytokine-STAT Signaling in Murine Systemic Lupus Erythematosus. PLoS ONE. 2009; 4(8):e6756. (Clone-specific: Flow cytometry). View Reference
Heim MH. The Jak-STAT pathway: specific signal transduction from the cell membrane to the nucleus. Eur J Clin Invest. 1996; 26(1):1-12. (Biology). View Reference
Hou J, Schindler U, Henzel WJ, Ho TC, Brasseur M, McKnight SL. An interleukin-4-induced transcription factor: IL-4 Stat. Science. 1994; 265(5179):1701-1706. (Biology).
Mikita T, Campbell D, Wu P, Williamson K, Schindler U. Requirements for interleukin-4-induced gene expression and functional characterization of Stat6. Mol Cell Biol. 1996; 16(10):5811-5820. (Biology).
Quelle FW, Shimoda K, Thierfelder W, et al. Cloning of murine Stat6 and human Stat6, Stat proteins that are tyrosine phosphorylated in responses to IL-4 and IL-3 but are not required for mitogenesis. Mol Cell Biol. 1995; 15(6):3336-3343. (Biology).

View All (8)

558243 Rev. 5

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.