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Two-color flow cytometric analysis of PLZF expression by human peripheral blood lymphocytes and mouse thymocytes. Left Panel: Human peripheral blood mononuclear cells 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 Alexa Fluor® 647 Mouse Anti-Human NCAM-1 (CD56) antibody (Cat. No. 563443) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-PLZF antibody (Cat. No. 564850; Right Plot). The two-color flow cytometric contour plots showing the correlated expression of CD56 versus PLZF (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Right Panel: BALB/c mouse thymocytes were stained with APC Hamster Anti-Mouse CD3e antibody (Cat. No. 553066/561826). The cells were washed and similarly fixed and permeabilized. The cells were then washed with BD Perm/Wash™ Buffer and stained with either PE Mouse IgG1, κ Isotype Control (Left Plot) or PE Mouse Anti-PLZF antibody (Right Plot). The two-color flow cytometric contour plots showing the correlated expression of CD3e versus PLZF (or Ig Isotype control staining) were derived from 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™ PE Mouse Anti-PLZF
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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.
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- 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.
- 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.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
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関連製品
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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