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Two color flow cytometric analysis of PLZF expression by mouse thymocytes and human peripheral blood mononuclear cells. Left Panel: Human peripheral blood mononuclear cells (PBMC) were fixed and permeabilized using BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were subsequently washed with BD Perm/Wash™ Buffer (Cat. No. 554723) and stained with PE Mouse Anti-Human NCAM-1 (CD56) (Cat. No. 563238) and Alexa Fluor® 647 Mouse Anti-PLZF antibodies. The two-color flow cytometric contour plots shows the coexpression of CD56 versus PLZF for gated events with the light-scatter characteristics of intact lymphocytes. Right Panel: BALB/c mouse thymocytes were stained with FITC Hamster Anti-Mouse CD3e antibody (Cat. No. 553061/553062/561827). The cells were washed and then fixed and permeabilized using BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were washed with BD Perm/Wash™ Buffer (Cat. No. 554723) and stained with Alexa Fluor® 647 Mouse Anti-PLZF antibody or matched isotype control (Cat. No. 557783/557732). The two-color flow cytometric contour plots shows the correlated expression patterns of CD3e versus PLZF for gated events with the forward and side light-scatter characteristics of intact thymocytes. Flow cytometric analyses were performed using a BD LSRFortessa™ Cell Analyzer System.
BD Pharmingen™ Alexa Fluor® 647 Mouse Anti-PLZF
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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
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- 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.
- 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
The R17-809 monoclonal antibody recognizes promyelocytic leukemia zinc finger protein (PLZF). PLZF is also known as zinc finger and BTB domain containing 16 (ZBTB16) and Zinc finger protein 145. PLZF is a member of the BTB/POZ-ZF family of transcription factors that includes Th-POK (Zbtb7b) which is involved in CD4+ T cell fate determination. PLZF serves multiple functions. In the immune system, PLZF is involved in the developmental regulation of innate immune lymphocytes including NKT cells, a subset of γδ T cells, and mucosal associated invariant T (MAIT) cells. The R17-809 hybridoma was generated from a mouse immunized with recombinant mouse PLZF protein. The R17-809 antibody crossreacts with both mouse and human PLZF.
Development References (3)
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Alonzo ES, Gottschalk RA, Das J, et al. Development of promyelocytic zinc finger and ThPOK-expressing innate gamma delta T cells is controlled by strength of TCR signaling and Id3. J Immunol. 2010; 184(3):1268-1279. (Biology). View Reference
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Alonzo ES, Sant'Angelo DB. Development of PLZF-expressing innate T cells. Curr Opin Immunol. 2011; 23(2):220-227. (Biology). View Reference
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Savage AK, Constantinides MG, Han J, et al. The transcription factor PLZF directs the effector program of the NKT cell lineage. Immunity. 2008; 29(3):391-403. (Biology). 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.
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.