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BD Phosflow™ PE Mouse anti-Stat3 (pS727)
Clone 49/p-Stat3 (RUO)
Analysis of Stat3 (pS727) in monocytes. Human peripheral blood mononuclear cells (PBMC) were either stimulated with 40 nM PMA (Sigma, P8139) for 15 minutes (shaded histogram) or unstimulated (open histogram). The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD PhosFlow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with PE Mouse anti-Stat3 (pS727, Cat. No. 558557). Monocytes were selected by scatter profile. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Analysis of Stat3 (pS727) in monocytes. Human peripheral blood mononuclear cells (PBMC) were either stimulated with 40 nM PMA (Sigma, P8139) for 15 minutes (shaded histogram) or unstimulated (open histogram). The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD PhosFlow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with PE Mouse anti-Stat3 (pS727, Cat. No. 558557). Monocytes were selected by scatter profile. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Product Details
BD Phosflow™
STAT3; STAT-3; APRF; Acute-phase response factor; HIES; ADMIO
Human (QC Testing), Mouse, Rat (Tested in Development)
Mouse IgG1
Phosphorylated Human Stat3
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_647232
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
558557 Rev. 2
Antibody Details
49/p-Stat3
The 49/p-Stat3 monoclonal antibody recognizes the S727-phosphorylated form of Signal transducer and activator of transcription 3 (Stat3/STAT3 isoform 1). The fluorochrome-conjugated formats have been evaluated using a human model system. However, the unconjugated form of this antibody is also effective for western blot analysis of human, mouse, and rat tissue. The Stat proteins function both as cytoplasmic signal transducers and as activators of transcription. Seven mammalian Stat proteins have been identified: Stat1-4, Stat5a, 5b, and Stat6. Stat3 is a 92-kDa protein that is activated as a DNA binding protein through cytokines, such as IL-6, and growth factors, such as EGF. Stat3 is phophorylated at serine 727 (S727) via the MAPK pathway. The S727 residue is located at a conserved Pro-X-Ser-Pro sequence, which is recognized by the protein kinase ERK. Activation through the S727 residue is thought to lead to initiation of transcription. Upon activation, Stat3 dimerizes, translocates to the nucleus, and binds DNA response elements thereby regulating gene expression. It appears that Stat3 binds to DNA as a homodimer, but it is also capable of binding as a heterodimer with Stat1. In addition to serine phosphorylation, Stat3 is also phosphorylated at tyrosine 705 by JAK1 in response to cytokine stimulation. Stat3 is widely expressed and can bind to the sis-inducible element (SIE) site from the c-fos promoter. This site is similar to the GAS element that is present in IFN--induced genes. Thus, phosphorylation of S727 in Stat3 occurs in response to growth factors and cytokines, and is essential for normal transcriptional activity.
558557 Rev. 2
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
496 nm, 566 nm
576 nm
558557 Rev.2
Citations & References
View product citations for antibody "558557" on CiteAb
Development References (4)
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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
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Imada K, Leonard WJ. The Jak-STAT pathway. Mol Immunol. 2000; 37:1-11. (Biology). View Reference
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Kanai M, Konda Y, Nakajima T, et al . Differentiation-inducing factor-1 (DIF-1) inhibits STAT3 activity involved in gastric cancer cell proliferation via MEK-ERK-dependent pathway. Oncogene. 2003; 22(22):548-554. (Biology). View Reference
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Schuringa JJ, Dekker LV, Vellenga E, Kruijer W. Sequential activation of Rac-1, SEK-1/MKK-4, and protein kinase Cδ is required for interleukin-6-induced STAT3 Ser-727 phosphorylation and transactivation. J Biol Chem. 2001; 276:27709-27715. (Biology).
558557 Rev. 2
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.