Multicolor flow cytometric analysis of SATB1 expressed in human peripheral blood lymphocytes. Panels 1a-c and 2a-c. Human peripheral blood mononuclear cells (PBMC) were surface stained with PE Mouse Anti-Human CD25 (Cat. No. 555432) and PerCP Mouse Anti-Human CD4 (Cat. No. 347324) antibodies followed by treatment with either the BD Cytofix/Cytoperm™ Fixation/Permeablization Kit (Cat. No. 554714, Panels 1a-c) or the Human FoxP3 Buffer Set (Cat. No. 560098, Panels 2a-c) as per the recommended protocols. The cells were then stained with Alexa Fluor® 647 Mouse Anti-Human SATB1 (Cat. No. 562378) and Alexa Fluor® 488 Mouse Anti-Human FoxP3 (Cat. No. 561181) antibodies. Events with the forward and side-light scatter characteristics of intact lymphocytes were reanalyzed for CD4 versus CD25 expression (Panels 1a and 2a) to gate for (1) CD25hiCD4+ (Treg), (2) CD25intCD4+, and (3) CD25negCD4+ T cell subsets as indicated. Panels 1b and 2b show overlaid SATB1 fluorescence histograms for the three gated CD4+ T cell subsets [ie, CD4 and CD25 gated subsets in Panels 1a and 2a (labeled 1-3) corresponding to histograms 1-3] using either the Cytofix/Cytoperm Buffer or FoxP3 Buffer, respectively. Panels 1c and 2c show the expression of SATB1 versus Foxp3 expression for the CD4+ T cell populations treated with Cytofix/Cytoperm Buffer or Foxp3 Buffer, respectively. Panels 1d,e and 2d,e. Human PBMC were surface stained with FITC Mouse Anti-Human CD4 (Cat. No. 555346) and PerCP-Cy™5.5 Mouse Anti-Human CD19 (Cat. No. 561295) antibodies followed by treatment with either Cytofix/Cytoperm Buffer (Cat. No. 554714, Panels 1d,e) or Human FoxP3 Buffer Set (Cat. No. 560098, Panels 2d,e) as per the recommended protocol. Cells were stained with A647 Mouse Anti-Human SATB1. Panels 1d,e and 2d,e show the expression of SATB1 in CD4+ T cell and CD19+ B cell populations. Flow cytometry was performed using a BD LSR™ II Flow Cytometer System.
Multicolor flow cytometric analysis of SATB1 expressed in human peripheral blood lymphocytes. Panels 1a-c and 2a-c. Human peripheral blood mononuclear cells (PBMC) were surface stained with PE Mouse Anti-Human CD25 (Cat. No. 555432) and PerCP Mouse Anti-Human CD4 (Cat. No. 347324) antibodies followed by treatment with either the BD Cytofix/Cytoperm™ Fixation/Permeablization Kit (Cat. No. 554714, Panels 1a-c) or the Human FoxP3 Buffer Set (Cat. No. 560098, Panels 2a-c) as per the recommended protocols. The cells were then stained with Alexa Fluor® 647 Mouse Anti-Human SATB1 (Cat. No. 562378) and Alexa Fluor® 488 Mouse Anti-Human FoxP3 (Cat. No. 561181) antibodies. Events with the forward and side-light scatter characteristics of intact lymphocytes were reanalyzed for CD4 versus CD25 expression (Panels 1a and 2a) to gate for (1) CD25hiCD4+ (Treg), (2) CD25intCD4+, and (3) CD25negCD4+ T cell subsets as indicated. Panels 1b and 2b show overlaid SATB1 fluorescence histograms for the three gated CD4+ T cell subsets [ie, CD4 and CD25 gated subsets in Panels 1a and 2a (labeled 1-3) corresponding to histograms 1-3] using either the Cytofix/Cytoperm Buffer or FoxP3 Buffer, respectively. Panels 1c and 2c show the expression of SATB1 versus Foxp3 expression for the CD4+ T cell populations treated with Cytofix/Cytoperm Buffer or Foxp3 Buffer, respectively. Panels 1d,e and 2d,e. Human PBMC were surface stained with FITC Mouse Anti-Human CD4 (Cat. No. 555346) and PerCP-Cy™5.5 Mouse Anti-Human CD19 (Cat. No. 561295) antibodies followed by treatment with either Cytofix/Cytoperm Buffer (Cat. No. 554714, Panels 1d,e) or Human FoxP3 Buffer Set (Cat. No. 560098, Panels 2d,e) as per the recommended protocol. Cells were stained with A647 Mouse Anti-Human SATB1. Panels 1d,e and 2d,e show the expression of SATB1 in CD4+ T cell and CD19+ B cell populations. Flow cytometry was performed using a BD LSR™ II Flow Cytometer System.
Product Details
BD Pharmingen™
SATB homeobox 1; Special AT-rich sequence-binding protein 1
Human (QC Testing), Mouse, Rat (Tested in Development)
Mouse IgG1
Human SATB1 aa. 550-667
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_11153310
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation/Permeabilization Kit RUO
Size
250 Tests
Cat No.
554714
Human FoxP3 Buffer Set RUO
Size
100 Tests
Cat No.
560098
PE Mouse Anti-Human CD25 RUO
Size
100 Tests
Cat No.
555432
FITC Mouse Anti-Human CD4 RUO
Size
100 Tests
Cat No.
555346
Alexa Fluor® 488 Mouse anti-Human FoxP3 RUO
Size
100 Tests
Cat No.
561181
562378 Rev. 2
Antibody Details
14/SATB1
The 14/SATB1 monoclonal antibody specifically binds to the special AT-rich sequence binding protein 1 that is encoded by the SATB1 (SATB homeobox 1) gene. Central to the formation of nucleosomes, which consist of octameric histone cores with defined segments of chromatin wound around them, are the interactions of nucleosomes with nuclear matrix components. Specific genomic DNA segments that interact with the nuclear matrix are called scaffold or matrix attachment regions (SARs or MARs). SATB1 is a homeodomain MAR binding protein. It recognizes ATC sequences that consist of stretches of A's, T's, and C's on one DNA strand. SATB1 is expressed by thymocytes (especially single positive CD4+ thymocytes), T and B lymphocytes, NK cells, and macrophages. SATB1 can recruit chromatin-remodeling factors and thereby regulate chromatin structure and gene expression. Recently it has been reported that repression of SATB1 was essential for the phenotype and function of mouse regulatory T cells but inhibitory for the establishment of transcriptional programs expressed by effector T cells.
562378 Rev. 2
Format Details
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 520-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.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
562378 Rev.2
Citations & References
View product citations for antibody "562378" on CiteAb
Development References (6)
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Alvarez JD, Yasui DH, Niida H, Joh T, Loh DY, Kohwi-Shigematsu T. The MAR-binding protein SATB1 orchestrates temporal and spatial expression of multiple genes during T-cell development. Genes Dev. 2000; 14(5):521-535. (Biology). View Reference
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Beyer M, Thabet Y, Müller R-U, et al. Repression of the genome organizer SATB1 in regulatory T cells is required for suppressive function and inhibition of effector differentiation. Nat Immunol. 2011; 12:898-907. (Clone-specific: Flow cytometry, Immunofluorescence, Western blot). View Reference
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Cai S, Kohwi-Shigematsu T.. Intranuclear relocalization of matrix binding sites during T cell activation detected by amplified fluorescence in situ hybridization.. 1999; 19(3):394-402. (Biology). View Reference
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Dickinson LA, Joh T, Kohwi Y, Kohwi-Shigematsu T. A tissue-specific MAR/SAR DNA-binding protein with unusual binding site recognition. Cell. 1992; 70(4):631-645. (Biology). View Reference
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Liu J, Bramblett D, Zhu Q. The matrix attachment region-binding protein SATB1 participates in negative regulation of tissue-specific gene expression. Mol Cell Biol. 1997; 17(9):5275-5287. (Biology). View Reference
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de Belle I, Cai S, Kohwi-Shigematsu T. The genomic sequences bound to special AT-rich sequence-binding protein 1 (SATB1) in vivo in Jurkat T cells are tightly associated with the nuclear matrix at the bases of the chromatin loops. J Cell Biol. 1998; 141(2):335-348. (Biology). View Reference
562378 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.