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BV421 Mouse Anti-Stat1 (pY701)
BV421 Mouse Anti-Stat1 (pY701)
Flow cytometric analysis of Stat1 (pY701) expression in human peripheral blood monocytes. Peripheral blood mononuclear cells (PBMC) were either left unstimulated (dashed line histogram) or stimulated (solid line histogram) with 100 ng/ml (final concentration) of Recombinant Human IFN-γ (Cat. No. 554617) for 15 minutes at 37°C. The PBMC were fixed (10 min, 37°C) with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then permeabilized (on ice, 30 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) and stained with BD Phosflow™ BV421 Mouse Anti-Stat1 (pY701) antibody (Cat. No. 562985). The fluorescence histograms 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 Stat1 (pY701) expression in human peripheral blood monocytes. Peripheral blood mononuclear cells (PBMC) were either left unstimulated (dashed line histogram) or stimulated (solid line histogram) with 100 ng/ml (final concentration) of Recombinant Human IFN-γ (Cat. No. 554617) for 15 minutes at 37°C. The PBMC were fixed (10 min, 37°C) with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and then permeabilized (on ice, 30 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) and stained with BD Phosflow™ BV421 Mouse Anti-Stat1 (pY701) antibody (Cat. No. 562985). The fluorescence histograms 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™
Signal Transducer and Activator of Transcription-1; STAT91; ISGF-3; CANDF7
Human (QC Testing), Mouse (Tested in Development), Rat (Predicted)
Mouse IgG2a
Phosphorylated Human Stat1 Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2737932
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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  8. Brilliant Violet™ 421 is a trademark of Sirigen.
  9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
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Antibody Details
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4a

Stat (Signal transducer and activators of transcription) proteins mediate the biological activity of cytokines, including interleukins, interferons, erythropoietin, and growth factors.  Ligand-receptor interaction activates constitutively-associated JAK family kinases as well as subsequent recruitment and activation of Stat proteins by tyrosine phosphorylation.  Active Stat proteins 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.  Stat1 and Stat2 are components of the ISGF3 (Interferon-Stimulated Gene Factor 3) complex, the primary transcription activator induced by interferon binding to a specific cell-surface receptor.  Stat1 has two alternatively spliced isoforms, 91-kDa Stat1α and 84-kDa Stat1β; Stat1α has 38 additional C-terminal amino acids.  In response to the binding of IFNα, IFNγ, EGF, PDGF, or CSF-1 to their respective receptors, the Stat1 subunits become tyrosine-phosphorylated at Y701, and the complex translocates to the nucleus. This forms an active complex that includes the DNA-binding p48 subunit, and is responsible for modulating interferon-stimulated genes (ISGs) transciption.

The 4a monoclonal antibody recognizes the phosphorylated Y701 in Stat1α and Stat1β.

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.

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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.
altImg
BV421
Violet 405 nm
407 nm
423 nm
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Citations & References
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Development References (6)

  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. Darnell JE Jr. STATs and gene regulation. Science. 1997; 277(5332):1630-1635. (Biology). View Reference
  3. Fu XY, Zhang JJ. Transcription factor p91 interacts with the epidermal growth factor receptor and mediates activation of the c-fos gene promoter. Cell. 1993; 74(6):1135-1145. (Biology). View Reference
  4. Perez OD, Mitchell D, Campos R, Gao GJ, Li L, Nolan GP. Multiparameter analysis of intracellular phosphoepitopes in immunophenotyped cell populations by flow cytometry. Curr Protoc Cytom. 2005; 6.20.1-6.20.22. (Clone-specific: Flow cytometry). View Reference
  5. Suni MA, Maino VC. Flow cytometric analysis of cell signaling proteins. Methods Mol Biol. 2011; 717:155-169. (Clone-specific). View Reference
  6. Tanaka S, Saito Y, Kunisawa J, et al. Development of mature and functional human myeloid subsets in hematopoietic stem cell-engrafted NOD/SCID/IL2rgammaKO mice. J Immunol. 2012; 188(12):6145-6155. (Clone-specific: Flow cytometry). View Reference
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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.