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Alexa Fluor™ 488 Mouse Anti-Nkx6.1
Alexa Fluor™ 488 Mouse Anti-Nkx6.1
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 Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 565572; dashed line histogram) or Alexa Fluor™ 488 Mouse Anti-Nkx6.1 antibody (Cat. No. 567608/567609; solid line histogram). Fluorescence histogram showing Nkx6.1 expression (or Ig Isotype staining) was derived from gated events with the side and forward light-scattering characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
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 Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 565572; dashed line histogram) or Alexa Fluor™ 488 Mouse Anti-Nkx6.1 antibody (Cat. No. 567608/567609; solid line histogram). Fluorescence histogram showing Nkx6.1 expression (or Ig Isotype staining) was derived from gated events with the side and forward light-scattering characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Product Details
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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
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 the dye under optimum conditions and unreacted dye was removed.

Recommended Assay Procedures

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.

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  6. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  11. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
567608 Rev. 2
Antibody Details
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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.

567608 Rev. 2
Format Details
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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
567608 Rev.2
Citations & References
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View product citations for antibody "567608" on CiteAb

Development References (6)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
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567608 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.