Analysis of PLK1 (pT210) in human epithelioid carcinoma. Hela S3 cells (ATCC CCL 2.2) were either stimulated with Nocodazole at 37°C for 12-16 hours (shaded histogram) or unstimulated (open histogram). The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 488 Mouse anti-PLK1 (pT210). Simultaneous staining with 7-AAD (Cat. No. 559925) demonstrates that the cells with upregulated PLK1 phosphorylation are in the G2/M phase of the cell cycle (data not shown). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
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Analysis of PLK1 (pT210) in human epithelioid carcinoma. Hela S3 cells (ATCC CCL 2.2) were either stimulated with Nocodazole at 37°C for 12-16 hours (shaded histogram) or unstimulated (open histogram). The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 488 Mouse anti-PLK1 (pT210). Simultaneous staining with 7-AAD (Cat. No. 559925) demonstrates that the cells with upregulated PLK1 phosphorylation are in the G2/M phase of the cell cycle (data not shown). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Product Details
BD Phosflow™
Human (QC Testing), Mouse (Predicted)
Mouse IgG1, κ
Phosphorylated Human PLK1 Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_647095
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® 488 under optimum conditions, and unreacted Alexa Fluor® 488 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).
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
558446 Rev. 2
Antibody Details
K50-483
Polo-like kinase (PLK1) is a serine/threonine kinase with structural similarities to Drosophila's Polo kinase and the Cdc5p of Saccharomyces cerevisiae. Like its invertebrate counterparts, PLK1 activity is required for DNA synthesis and is regulated throughout the cell cycle. Furthermore, PLK1 is highly expressed in primary tumors. It associates with the mitotic spindle during mitosis suggesting that, in addition to its role during S phase, PLK1 may play a role during chromosome segregation. This is consistent with its potential role in cancer development. Threonine 210 (T210) is one of the major phoshorylation sites in activated PLK1 obtained from human mitotic cells.
The K50-483 monoclonal antibody recognizes the phosphorylated T210 of human PLK1.
558446 Rev. 2
Format Details
Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
558446 Rev.2
Citations & References
View product citations for antibody "558446" on CiteAb
Development References (4)
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Ando K, Ozaki T, Yamamoto H, et al. Polo-like kinase 1 (Plk1) inhibits p53 function by physical interaction and phosphorylation. J Biol Chem. 2004; 279(24):25549-25561. (Biology).
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Jang Y-J, Ma S, Terada Y, Erikson RL. Phosphorylation of threonine 210 and the role of serine 137 in the regulation of mammalian Polo-like kinase. J Biol Chem. 2002; 277(46):44115-44120. (Biology).
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Peter B, Gleixner K, Cerny-Reiterer S, et al. Polo-like kinase-1 as a novel target in neoplastic mast cells: demonstration of growth-inhibitory effects of small interfering RNA and the polo-like kinase-1 targeting drug BI 2536. Haematologica. 2011; 96(5):672-680. (Clone-specific: Immunocytochemistry (cytospins)). View Reference
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Yamashiro S, Yamakita Y, Totsukawa G, et al. Myosin phosphatase targeting subunit1 regulates mitosis by antagonizing polo-like kinase1. Dev Cell. 2008; 14(5):787-797. (Clone-specific: Immunofluorescence, Western blot). View Reference
558446 Rev. 2
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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.