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BV421 Mouse Anti-Stat3 (pS727)
BV421 Mouse Anti-Stat3 (pS727)

Flow cytometric analysis of Stat3 (pS727) expression in human monocytes. Human peripheral blood mononuclear cells (PBMC) were either left untreated (dashed line histogram) or stimulated (solid line histogram) with 40 nM PMA (Phorbol 12-Myristate 13-Acetate; Sigma Cat. No. P8139; 15 min at 37°C). The cells were fixed (10 min) with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized (30 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050).  The cells were then stained with BD Phosflow™ BV421 Mouse Anti-Stat3 (pS727) antibody (Cat. No. 565416). The fluorescence histograms showing Stat3 (pS727) expression were derived from gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of Stat3 (pS727) expression in human monocytes. Human peripheral blood mononuclear cells (PBMC) were either left untreated (dashed line histogram) or stimulated (solid line histogram) with 40 nM PMA (Phorbol 12-Myristate 13-Acetate; Sigma Cat. No. P8139; 15 min at 37°C). The cells were fixed (10 min) with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized (30 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050).  The cells were then stained with BD Phosflow™ BV421 Mouse Anti-Stat3 (pS727) antibody (Cat. No. 565416). The fluorescence histograms showing Stat3 (pS727) expression were derived from gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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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)
5 µl
AB_2739226
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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565416 Rev. 1
Antibody Details
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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.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

565416 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
565416 Rev.1
Citations & References
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Development References (5)

  1. 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
  2. Imada K, Leonard WJ. The Jak-STAT pathway. Mol Immunol. 2000; 37:1-11. (Biology). View Reference
  3. Iwata Y, Furuichi K, Hashimoto S, et al. Pro-inflammatory/Th1 gene expression shift in high glucose stimulated mesangial cells and tubular epithelial cells. Biochem Biophys Res Commun. 2014; 443(3):969-974. (Clone-specific: Flow cytometry). View Reference
  4. 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. (Clone-specific: Western blot). View Reference
  5. Lu SX, Alpdogan O, Lin J, et al. STAT-3 and ERK 1/2 phosphorylation are critical for T-cell alloactivation and graft-versus-host disease. Blood. 2008; 112(13):5254-5258. (Clone-specific: Flow cytometry). View Reference
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565416 Rev. 1

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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.