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
View product citations for antibody "563022" on CiteAb
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
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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.