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BV421 Rat Anti-Mouse Notch1
BV421 Rat Anti-Mouse Notch1

Flow cytometric analysis using BD OptiBuild™ BV421 Rat Anti-Mouse Notch1 antibody (Cat. No. 744812; solid line histogram) on BALB/c mouse spleen, with Isotype Control (dotted line histogram). Flow cytometry was performed using a BD LSRFortessa™ Flow Cytometer System.

Flow cytometric analysis using BD OptiBuild™ BV421 Rat Anti-Mouse Notch1 antibody (Cat. No. 744812; solid line histogram) on BALB/c mouse spleen, with Isotype Control (dotted line histogram). Flow cytometry was performed using a BD LSRFortessa™ Flow Cytometer System.

Product Details
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BD OptiBuild™
Notch1; Notch 1; NOTC1; lin-12; Mis6; Motch A; mT14; N1; NICD; p300; Tan1
Mouse (Tested in Development)
Rat IgG2a, κ
Mouse Notch1 extracellular domain Recombinant Protein
Flow cytometry (Qualified)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.

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 BD Horizon BV421 under optimal conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. This antibody was developed for use in flow cytometry.
  2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  3. Researchers should determine the optimal concentration of this reagent for their individual applications.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at
  7. Please refer to for technical protocols.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  10. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
744812 Rev. 2
Antibody Details
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The 22E5.5 monoclonal antibody specifically binds to the extracellular domain of mouse Notch1. This Type 1 transmembrane protein is a member of the Notch family that includes Notch1-4. Notch ligands include membrane-bound Jagged1, Jagged2 and Delta-like1, 3 and 4. Notch receptors mediate juxtacrine signals between adjacent cells. Upon ligand binding, Notch1 undergoes proteolytic cleavage that results in the release of the Notch intracellular domain, NICD. NICD translocates to the nucleus where it forms a transcriptional activator complex with the RBP-J DNA binding factor and other transcriptional cofactors. These multimeric complexes regulate the expression of multiple genes including those that orchestrate many facets of embryonic development and the subsequent functioning of multiple organ systems such as the immune and nervous systems. Within the immune system, Notch signaling significantly affects the differentiation and survival of numerous cell types including thymocytes and subsets of T and B lymphocytes and dendritic cells. In altered forms, Notch1 has been implicated in transformation or progression in some T-cell neoplasms. The mN1A monoclonal antibody specifically binds to an intracellular domain of mouse Notch1.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

744812 Rev. 2
Format Details
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The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Violet 405 nm
407 nm
423 nm
744812 Rev.2
Citations & References
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Development References (7)

  1. Anderson AC, Robey EA, Huang YH. Notch signaling in lymphocyte development. Curr Opin Genet Dev. 2001; 11(5):554-560. (Biology). View Reference
  2. Artavanis-Tsakonas S, Rand MD, Lake RJ. Notch signaling: cell fate control and signal integration in development.. Science. 1999; 284(5415):770-6. (Biology). View Reference
  3. Fiorini E, Merck E, Wilson A, et al. Dynamic regulation of Notch1 and Notch2 surface expression during T cell development and activation revealed by novel monoclonal antibodies. J Immunol. 2009; 183(11):7212-7222. (Immunogen: Flow cytometry, Immunofluorescence). View Reference
  4. Helbig C, Gentek R, Backer RA, et al. Notch controls the magnitude of T helper cell responses by promoting cellular longevity. Proc Natl Acad Sci U S A. 109(23):9041-9046. (Biology). View Reference
  5. Kojika S, Griffin JD. Notch receptors and hematopoiesis. Exp Hematol. 2001; 29(9):1041-1052. (Biology). View Reference
  6. Radtke F, Fasnacht N, Macdonald HR. Notch signaling in the immune system. Immunity. 2010; 32(1):14-27. (Biology). View Reference
  7. Walker L, Carlson A, Tan-Pertel HT, Weinmaster G, Gasson J. The notch receptor and its ligands are selectively expressed during hematopoietic development in the mouse. Stem Cells. 2001; 19(6):543-552. (Biology). View Reference
View All (7) View Less
744812 Rev. 2

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.