Western Blot analysis of Nkx6.1 in mouse pancreatic tumor (insulinoma) cell lysate. Lysate from Beta-T-C6 cells (ATCC, CRL-11506™) was probed with Purified Mouse Anti-Nkx6.1 monoclonal antibody at titrations of 0.5 (lane 1), 0.25 (lane 2), and 0.125 μg/ml (lane 3). Nkx6.1 is identified as a doublet of approximately 44 kDa.
Immunoflourescent staining of Nkx6.1 in mouse pancreatic tumor (insulinoma) cells. Beta-T-C6 cells (ATCC, CRL-11506™) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm III Buffer (Cat. No. 558050, and stained with Purified Mouse Anti-Nkx6.1 monoclonal antibody (pseudo-colored green) at 1.2 μg/mL. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Life Technologies), and counter-staining was with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD AttoVision™ Software.
Western Blot analysis of Nkx6.1 in mouse pancreatic tumor (insulinoma) cell lysate. Lysate from Beta-T-C6 cells (ATCC, CRL-11506™) was probed with Purified Mouse Anti-Nkx6.1 monoclonal antibody at titrations of 0.5 (lane 1), 0.25 (lane 2), and 0.125 μg/ml (lane 3). Nkx6.1 is identified as a doublet of approximately 44 kDa.
Immunoflourescent staining of Nkx6.1 in mouse pancreatic tumor (insulinoma) cells. Beta-T-C6 cells (ATCC, CRL-11506™) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm III Buffer (Cat. No. 558050, and stained with Purified Mouse Anti-Nkx6.1 monoclonal antibody (pseudo-colored green) at 1.2 μg/mL. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Life Technologies), and counter-staining was with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD AttoVision™ Software.
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
Western blot (Routinely Tested), Bioimaging, Immunofluorescence, Intracellular staining (flow cytometry) (Tested During Development)
~44 kDa
0.5 mg/ml
AB_2737958
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- All other brands are trademarks of their respective owners.
- 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.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
HRP Goat Anti-Mouse Ig RUO
Size
1 mL
Cat No.
554002
563022 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.
563022 Rev. 1
Format Details
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
563022 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
563022 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.
