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Analysis of PTEN expression in HeLa S3 cells. Cells from the human HeLa S3 (Cervical adenocarcinoma, ATCC CCL-2.2) cell line were fixed (10 min, 37°C) with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, and then stained with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; dashed line histogram) or BD Horizon BUV395 Mouse Anti-PTEN antibody (Cat. No. 566638/566639; solid line histogram) at 0.5 µg/ml. The fluorescence histogram showing PTEN expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact HeLa S3 cells. Data shown in this Technical Data Sheet are not lot specific.
BD Phosflow™ BUV395 Mouse Anti-PTEN
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD Optibuild Brilliant reagents) are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- 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
Cancer can develop when cells escape normal growth control mechanisms through mutations in proto-oncogenes or tumor suppressor genes. A characteristic of most oncogene and tumor suppressor gene products is that they are components of signal transduction pathways that are essential for maintaining cellular homeostasis. PTEN (phosphatase and tensin homolog), also known as MMAC1 (mutated in multiple advanced cancers 1), is a tumor suppressor gene that is mutated at high frequency in multiple tumor types. The protein encoded by PTEN is a phosphatase that preferentially dephosphorylates phosphoinositide substrates. It is believed that a mechanism by which PTEN mutations cause tumors is the loss of its negative control on the phosphoinositide 3-kinase signaling pathway that regulates cell growth and survival. PTEN also plays a role in the maintenance of hematopoietic stem cells.
The A2B1 monoclonal antibody recognizes PTEN, regardless of phosphorylation status.
The antibody was conjugated to BD Horizon BUV395 which has been exclusively developed by BD Biosciences as an optimal dye for use on a 355 nm laser equipped instrument. With an Ex Max at 348 nm and an Em Max at 395 nm, this dye has virtually no spillover into any other detector. BD Horizon BUV395 can be excited with a 355 nm laser and detected with a 379/28 filter.
Development References (8)
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Cantley LC, Neel BG. New insights into tumor suppression: PTEN suppresses tumor formation by restraining the phosphoinositide 3-kinase/AKT pathway. Proc Natl Acad Sci U S A. 1999; 96(8):4240-4245. (Biology). View Reference
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Goberdhan DCI, Wilson C. PTEN: tumour suppressor, multifunctional growth regulator and more. Hum Mol Genet. 2003; 12(2):R238-R248. (Biology).
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Levine RA, Forest T, Smith C. Tumor suppressor PTEN is mutated in canine osteosarcoma cell lines and tumors. Vet Pathol. 2002; 39:372-378. (Clone-specific: Immunohistochemistry).
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Raftopoulou M, Etienne-Manneville S, Self A, Nicholls S, Hall A. Regulation of cell migration by the C2 domain of the tumor suppressor PTEN. Science. 2004; 303:1179-1181. (Biology).
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Steck PA, Pershouse MA, Jasser SA, et al. Identification of a candidate tumour suppressor gene, MMAC1 at chromosome 10q23.3 that is mutated in multiple advanced cancers. Nat Genet. 1997; 15(4):356-362. (Biology). View Reference
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Wu R-C, Li X, Schönthal AH. Transcriptional activation of p21WAF1 by PTEN/MMAC1 tumor suppressor. Mol Cell Biochem. 2000; 203:59-71. (Immunogen: Western blot).
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Yilmaz OH, Valez R, Theisen BK, et al. Pten dependence distinguishes haematopoietic stem cells from leukaemia-initiating cells. Nature. 2006; 441:475-482. (Biology).
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Zhang J, Grindley JC, Yin T, et al. PTEN maintains haematopoietic stem cells and acts in lineage choice and leukaemia prevention. Nature. 2006; 441:518-522. (Biology).
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.