Skip to main content Skip to navigation
Hamburger Menu
Search icon
Locaation Selector Locaation Selector
New Zealand (English)
Profile Icon Profile Icon
Sign-in/Register UserName
Products

Link Arrow
Solutions

Link Arrow
Discover & Learn

Link Arrow
Resources & Tools

Link Arrow
Support

Link Arrow
Close Menu
Website Feedback
Alert Icon
S38316_612597_image1_rev.png

Flow cytometric analysis of Stat1 (pY701).  U-937 cells (Human histiocytic lymphoma; ATCC CRL-1593.2) were either left unstimulated (unshaded) or stimulated (shaded) with 1000 U/mL recombinant human IFN-γ (Cat. No. 554617) for 15 minutes at 37°C. Cells were fixed with BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37°C and then permeabilized by adding BD Phosflow™ Perm Buffer III (Cat. No. 558050 for 30 minutes on ice. Cells were then washed twice in BD Pharmingen™ Stain Buffer (Cat. No. 554656) and stained with the Alexa Fluor® 647 mouse anti-Stat1 (pY701) antibody. Cells were analyzed on a BD FACSCalibur™ flow cytometery instrument. For intracellular staining of human whole blood, BD Phosflow™ Lyse/Fix buffer (Cat. No. 558049) may be used for fixation.

S38316_612597_image1_rev.png

Flow cytometric analysis of Stat1 (pY701).  U-937 cells (Human histiocytic lymphoma; ATCC CRL-1593.2) were either left unstimulated (unshaded) or stimulated (shaded) with 1000 U/mL recombinant human IFN-γ (Cat. No. 554617) for 15 minutes at 37°C. Cells were fixed with BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37°C and then permeabilized by adding BD Phosflow™ Perm Buffer III (Cat. No. 558050 for 30 minutes on ice. Cells were then washed twice in BD Pharmingen™ Stain Buffer (Cat. No. 554656) and stained with the Alexa Fluor® 647 mouse anti-Stat1 (pY701) antibody. Cells were analyzed on a BD FACSCalibur™ flow cytometery instrument. For intracellular staining of human whole blood, BD Phosflow™ Lyse/Fix buffer (Cat. No. 558049) may be used for fixation.

S38316_612597_image1_rev.png S38316_612597_image1_rev.png Red-Cap-Red-Label.jpg

Flow cytometric analysis of Stat1 (pY701).  U-937 cells (Human histiocytic lymphoma; ATCC CRL-1593.2) were either left unstimulated (unshaded) or stimulated (shaded) with 1000 U/mL recombinant human IFN-γ (Cat. No. 554617) for 15 minutes at 37°C. Cells were fixed with BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37°C and then permeabilized by adding BD Phosflow™ Perm Buffer III (Cat. No. 558050 for 30 minutes on ice. Cells were then washed twice in BD Pharmingen™ Stain Buffer (Cat. No. 554656) and stained with the Alexa Fluor® 647 mouse anti-Stat1 (pY701) antibody. Cells were analyzed on a BD FACSCalibur™ flow cytometery instrument. For intracellular staining of human whole blood, BD Phosflow™ Lyse/Fix buffer (Cat. No. 558049) may be used for fixation.

Flow cytometric analysis of Stat1 (pY701).  U-937 cells (Human histiocytic lymphoma; ATCC CRL-1593.2) were either left unstimulated (unshaded) or stimulated (shaded) with 1000 U/mL recombinant human IFN-γ (Cat. No. 554617) for 15 minutes at 37°C. Cells were fixed with BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37°C and then permeabilized by adding BD Phosflow™ Perm Buffer III (Cat. No. 558050 for 30 minutes on ice. Cells were then washed twice in BD Pharmingen™ Stain Buffer (Cat. No. 554656) and stained with the Alexa Fluor® 647 mouse anti-Stat1 (pY701) antibody. Cells were analyzed on a BD FACSCalibur™ flow cytometery instrument. For intracellular staining of human whole blood, BD Phosflow™ Lyse/Fix buffer (Cat. No. 558049) may be used for fixation.

Product Details
Down Arrow Up Arrow


BD Phosflow™
Signal Transducer and Activator of Transcription-1; STAT91; ISGF-3; CANDF7
Human (QC Testing), Mouse (Tested in Development)
Mouse IgG2a
Phosphorylated Human Stat1 Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_399880
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 to the dye under optimum conditions and unreacted dye was removed.
Show More blue down arrow Show Less blue up arrow

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Show More blue down arrow Show Less blue up arrow

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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  7. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Show More blue down arrow Show Less blue up arrow
612597 Rev. 7
Antibody Details
Down Arrow Up Arrow
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.

612597 Rev. 7
Format Details
Down Arrow Up Arrow
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
Alexa Fluor™ 647
653 nm
669 nm
612597 Rev.7
Citations & References
Down Arrow Up Arrow
View product citations for antibody "612597" on CiteAb

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
View All (6) View Less
612597 Rev. 7

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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.