Flow cytometric analysis of NeuroD1 expression on human retinoblastoma. Y79 cells (ATCC, HTB-18™) 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® 647 Mouse IgG1, κ isotype control (dashed line, Cat. No. 557732) or Alexa Fluor® 647 Mouse anti-NeuroD1 monoclonal antibody at matched concentrations. Flow cytometry was performed on a BD CantoII™ flow cytometry system. We do not recommend BD Phosflow™ Perm/Wash Buffer I for permeabilization.
Flow cytometric analysis of NeuroD1 expression on human retinoblastoma. Y79 cells (ATCC, HTB-18™) 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® 647 Mouse IgG1, κ isotype control (dashed line, Cat. No. 557732) or Alexa Fluor® 647 Mouse anti-NeuroD1 monoclonal antibody at matched concentrations. Flow cytometry was performed on a BD CantoII™ flow cytometry system. We do not recommend BD Phosflow™ Perm/Wash Buffer I for permeabilization.
Product Details
BD Pharmingen™
Class A basic helix-loop-helix protein 3, bHLHa3, BETA2, NeuroD
Human (QC Testing), Mouse (Tested in Development)
Mouse IgG1, κ
Human NeuroD1 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested), Bioimaging, Immunofluorescence (Not Recommended)
5 µl
AB_2738279
Aqueous buffered solution containing BSA, protein stabilizer, 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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was 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).
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- All other brands are trademarks of their respective owners.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557732
Purified Mouse Anti-NeuroD1 RUO
Size
0.1 mg
Cat No.
563000
563566 Rev. 1
Antibody Details
R8-294
The R8-294 monoclonal antibody specifically binds to Neurogenic differentiation factor 1 (NeuroD1). NeuroD1 is a basic helix-loop-helix (bHLH) transcription factor that is also known as BHLHA3, Beta-cell E-box transactivator 2, BETA2 and NeuroD. NeuroD1 interacts with other bHLH proteins and binds the insulin E-box sequence to regulate pancreatic islet cell development. It also plays a key role in central nervous system (CNS) and sensory nervous system neuron development. Specifically, it is required for the terminal differentiation of neurons during late stages of neurogenesis in the CNS (cerebral cortex, hippocampus, and cerebellum) and the sensory nervous system (eye, inner ear, and olfactory system). During antibody development, the purified R8-294 monoclonal antibody was found to detect NeuroD1 by western blot and indirect immunofluorescent staining followed by flow cytometric analysis or imaging analysis.
563566 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
563566 Rev.1
Citations & References
Development References (9)
-
Cherry TJ, Wang S, Bormuth I, Schwab M, Olson J, Cepko CL. NeuroD factors regulate cell fate and neurite stratification in the developing retina. J Neurosci. 2011; 31(20):7365-7379. (Biology). View Reference
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Cho JH, Tsai MJ. The role of BETA2/NeuroD1 in the development of the nervous system. Mol Neurobiol. 2004; 30(1):35-47. (Biology). View Reference
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Gao Z, Ure K, Ables JL, et al. Neurod1 is essential for the survival and maturation of adult-born neurons. Nat Neurosci. 2009; 12(9):1090-1092. (Biology). View Reference
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Habener JF, Kemp DM, Thomas MK. Minireview: transcriptional regulation in pancreatic development. Endocrinology. 2005; 146(3):1025-1034. (Biology). View Reference
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Kuwabara T, Hsieh J, Muotri A, et al. Wnt-mediated activation of NeuroD1 and retro-elements during adult neurogenesis. Nat Neurosci. 2009; 12(9):1097-10105. (Biology). View Reference
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Naya FJ, Huang HP, Qiu Y, et al. Diabetes, defective pancreatic morphogenesis, and abnormal enteroendocrine differentiation in BETA2/neuroD-deficient mice. Genes Dev. 1997; 11(18):2323-2334. (Biology). View Reference
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Naya FJ, Stellrecht CM, Tsai MJ. Tissue-specific regulation of the insulin gene by a novel basic helix-loop-helix transcription factor. Genes Dev. 1995; 9(8):1009-1019. (Biology). View Reference
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Pang ZP, Yang N, Vierbuchen T, et al. Induction of human neuronal cells by defined transcription factors. Nature. 2011; 476(7359):220-223. (Biology). View Reference
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Tamimi R, Steingrimsson E, Copeland NG, et al. The NEUROD gene maps to human chromosome 2q32 and mouse chromosome 2. Genomics. 1996; 15(34):418-421. (Biology). View Reference
563566 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.
