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      PE Mouse Anti-Nkx2.2
      PE Mouse Anti-Nkx2.2
      Flow cytometric analysis of Nkx2.2 expression in human embryonic kidney cells. 293-F cells (Life Technologies R790-07) were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574). Fixed cells were then stained with either PE Mouse IgG2b, κ Isotype Control (Cat. No. 555058, dashed line) or PE Mouse Anti-Nkx2.2 (solid line). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact 293-F cells. Flow cytometry was performed using a BD FACSCanto™ II Flow Cytometer System.
      PE Mouse Anti-Nkx2.2
      Flow cytometric analysis of Nkx2.2 expression in human embryonic kidney cells. 293-F cells (Life Technologies R790-07) were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574). Fixed cells were then stained with either PE Mouse IgG2b, κ Isotype Control (Cat. No. 555058, dashed line) or PE Mouse Anti-Nkx2.2 (solid line). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact 293-F cells. Flow cytometry was performed using a BD FACSCanto™ II Flow Cytometer System.
      Product Details
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      BD Pharmingen™
      NKX2-2; NKX2B; Nkx2-2; Nkx-2.2; NK-2 homolog B; glycoprotein GP330
      Human (QC Testing), Mouse (Tested in Development), Rat, Chicken (Reported)
      Mouse BALB/c IgG2b, κ
      Chicken Nkx2.2 Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      AB_2738923
      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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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      Recommended Assay Procedures

      PE Mouse Anti-Nkx2.2 effectively stains 293-F cells when using either the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574) or BD Cytofix™ Fixation Buffer (Cat. No. 554655) with BD Phosflow™ Perm Buffer III (Cat. No. 558050).

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      Product Notices

      1. 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).
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      5. 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.
      6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      564730 Rev. 1
      Antibody Details
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      74.5A5

      The 74.5A5 monoclonal antibody specifically binds to Homeobox protein Nkx-2.2 (Nkx2.2), which is a transcription factor required for the differentiation of ventral neuronal populations in the central nervous system and endocrine cell populations in the pancreas and intestine. Within the pancreas, Nkx2.2 is expressed in many cell subsets and plays important roles in islet and β-cell differentiation and maturation. Mice with a homozygous disruption of the Nkx2-2 gene lack mature β cells and die shortly after birth with severe hyperglycemia. Similarly to the Nkx2.2-ablated mice, homozygous mutations in the human NKX2-2 gene have been described in diabetic patients and associated with the etiology of neonatal diabetes. The NKX2-2 gene is also a target of the Ewing sarcoma specific protein EWS-FLI. Nkx2.2 expression can be monitored in order to determine cell fate changes in cultures for the generation of human pluripotent stem cell-derived insulin-producing cells as well as oligodendrocyte progenitors.

      564730 Rev. 1
      Format Details
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      PE
      R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      564730 Rev.1
      Citations & References
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      View product citations for antibody "564730" on CiteAb

      Development References (6)

      1. Briscoe J, Sussel L, Serup P, et al. Homeobox gene Nkx2.2 and specification of neuronal identity by graded Sonic hedgehog signalling.. Nature. 1999; 398(6728):622-7. (Clone-specific: Immunohistochemistry). View Reference
      2. D'Amour KA, Bang AG, Eliazer S, et al . Production of pancreatic hormone-expressing endocrine cells from human embryonic stem cells. Nat Biotechnol. 2006; 24(12):1481-1483. (Clone-specific: Immunofluorescence, Western blot). View Reference
      3. Ericson J, Rashbass P, Schedl A, et al. Pax6 controls progenitor cell identity and neuronal fate in response to graded Shh signalling. Cell. 1997; 90(1):169-180. (Immunogen: Immunohistochemistry). View Reference
      4. Sussel L, Kalamaras J, Hartigan-O'Connor DJ, et al. Mice Lacking the homeodomain transcription factor Nkx2.2 have diabetes due to arrested differentiation of pancreatic beta cells. Development. 1998; 125(12):2213-2221. (Clone-specific: Immunohistochemistry). View Reference
      5. Wang S, Bates J, Li X, et al. Human iPSC-derived oligodendrocyte progenitor cells can myelinate and rescue a mouse model of congenital hypomyelination. Cell Stem Cell. 2013; 12(2):252-264. (Clone-specific: Immunofluorescence). View Reference
      6. Yoshida A, Sekine S, Tsuta K, Fukayama M, Furuta K, Tsuda H. NKX2.2 is a useful immunohistochemical marker for Ewing sarcoma. Am J Surg Pathol. 2012; 36(7):993-999. (Biology). View Reference
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      564730 Rev. 1

       

      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.