Flow cytometric analysis of Nkx6.1 expression in mouse pancreatic tumor (insulinoma) cells. Beta-T-C6 (ATCC, CRL-11506™) cells were fixed with BD Cytofix™ fixation buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm buffer III (Cat. No. 558050). The cells were stained with either PE Mouse IgG1, κ isotype control (dashed line, Cat. No. 554680) or PE Mouse Anti-Nkx6.1 monoclonal antibody (solid line) at matched concentrations. Flow cytometry was performed on a BD LSRFortessa™ flow cytometry system.
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Flow cytometric analysis of Nkx6.1 expression in mouse pancreatic tumor (insulinoma) cells. Beta-T-C6 (ATCC, CRL-11506™) cells were fixed with BD Cytofix™ fixation buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm buffer III (Cat. No. 558050). The cells were stained with either PE Mouse IgG1, κ isotype control (dashed line, Cat. No. 554680) or PE Mouse Anti-Nkx6.1 monoclonal antibody (solid line) at matched concentrations. Flow cytometry was performed on a BD LSRFortessa™ flow cytometry system.
Product Details
BD Pharmingen™
Nkx6.1, NKX6-1, NKX6A, Nkx6-1, Nkx6.1, Nkx6a
Mouse (QC Testing), Human (Tested in Development)
Mouse IgG1, κ
Human Nkx6.1 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2716792
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.
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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- All other brands are trademarks of their respective owners.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
PE Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
554680
563023 Rev. 1
Antibody Details
R11-560
The R11-560 monoclonal antibody specifically binds to human and mouse homeobox protein Nkx6.1. The transcription factor Nkx6.1 plays a role in pancreatic islet beta cell development through the regulation of HNF1a and insulin genes. Nkx6.1 is considered to be a highly specific transcription factor marking pancreatic beta islet cells, where it is expressed in both developing and mature cells. In addition, Nkx6.1 functions as a transcriptional repressor induced by the Sonic Hedgehog (Shh) signaling pathway in fate determination of several neuronal cell types. During antibody development, the purified R11-560 monoclonal antibody was found to detect Nkx6.1 by western blot and indirect immunofluorescent staining followed by flow cytometric analysis or imaging analysis.
563023 Rev. 1
Format Details
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.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
563023 Rev.1
Citations & References
Development References (6)
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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. (Biology). View Reference
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Donelan W, Koya V, Li SW, Yang LJ. Distinct regulation of hepatic nuclear factor 1alpha by NKX6.1 in pancreatic beta cells. J Biol Chem. 2010; 285(16):12181-12189. (Biology). View Reference
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Inoue H, Rudnick A, German MS, Veile R, Donis-Keller H, Permutt MA. Isolation, characterization, and chromosomal mapping of the human Nkx6.1 gene (NKX6A), a new pancreatic islet homeobox gene. Genomics. 1997; 40:367-370. (Biology). View Reference
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Nelson SB, Schaffer AE, Sander M. The transcription factors Nkx6.1 and Nkx6.2 possess equivalent activities in promoting beta-cell fate specification in Pdx1+ pancreatic progenitor cells. Development. 2007; 134(13):2491-2500. (Biology). View Reference
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Pedersen IL, Klinck R, Hecksher-Sorensen J, et al. Generation and characterization of monoclonal antibodies against the transcription factor Nkx6.1. J Histochem Cytochem. 2006; 54(5):567-574. (Biology). View Reference
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Prakash N, Puelles E, Freude K, et. al. Nkx6-1 controls the identity and fate of red nucleus and oculomotor neurons in the mouse midbrain. Development. 2009; 136(15):2545-2555. (Biology). View Reference
563023 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.
