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PE Mouse Anti-Mouse Notch1
PE Mouse Anti-Mouse Notch1

Expression of Notch1 in double-negative thymocytes. Fixed and permeabilized C57BL/6 thymocytes were stained with either PE Mouse anti-Mouse Notch1 or PE Mouse IgG1, κ Isotype Control MOPC-31C (Cat. No. 550617, open histogram). Cells were then stained with FITC Rat anti-Mouse CD4 (Cat. No. 553046/553047) and APC Rat anti-Mouse CD8a (Cat. No. 553035). For analysis of Notch1 expression, an electronic gate was set to include only the CD4-CD8a subpopulation.  Flow cytometry was performed on a BD FACSCalibur™ Flow Cytometry System.

Expression of Notch1 in double-negative thymocytes. Fixed and permeabilized C57BL/6 thymocytes were stained with either PE Mouse anti-Mouse Notch1 or PE Mouse IgG1, κ Isotype Control MOPC-31C (Cat. No. 550617, open histogram). Cells were then stained with FITC Rat anti-Mouse CD4 (Cat. No. 553046/553047) and APC Rat anti-Mouse CD8a (Cat. No. 553035). For analysis of Notch1 expression, an electronic gate was set to include only the CD4-CD8a subpopulation.  Flow cytometry was performed on a BD FACSCalibur™ Flow Cytometry System.

Product Details
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BD Pharmingen™
Notch1; Notch 1; NOTC1; lin-12; Mis6; Motch A; mT14; N1; NICD; p300; Tan1
Mouse (QC Testing), Human (Reported)
Mouse IgG1, κ
Mouse Notch1 GST fusion protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_394454
Aqueous buffered solution containing ≤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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
552768 Rev. 2
Antibody Details
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mN1A

The Notch family of transmembrane receptors controls cell-fate "decisions" during the development of many organs in a wide variety of animal species. After binding its ligand, a Notch receptor is cleaved in its transmembrane domain, and the resulting intracellular domain dissociates from the membrane and translocates to the nucleus, where it is able to suppress the expression of lineage-specific genes by interacting with transcriptional repressors. The mN1A antibody reacts with the intracellular domain of mouse and human Notch1, but not with mouse Notch2, 3, or 4. In the mouse, Notch1 mRNA is expressed in mouse hematopoietic cells of the fetal liver and adult thymus and bone marrow. Using mAb mN1A, Notch1 is detected in CD4-CD8- (double-negative) and CD4-CD8+ thymocytes. Studies of Notch1-transgenic cells and Notch1-null mice indicate that the receptor is involved in the regulation of lymphopoiesis and myelopoiesis. The mN1A mAb does not cross-react with rat thymocytes.  An alternative anti-mouse Notch1 monoclonal antibody, clone 22E5.5, specifically binds to an extracellular domain of mouse Notch1.

552768 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
552768 Rev.2
Citations & References
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Development References (8)

  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. Huppert SS, Le A, Schroeter EH, et al. Embryonic lethality in mice homozygous for a processing-deficient allele of Notch1. Nature. 2000; 405(6789):966-970. (Immunogen). View Reference
  4. Kawamata S, Du C, Li K, Lavau C. Notch1 perturbation of hemopoiesis involves non-cell- autonomous modifications. J Immunol. 2002; 168(4):1738-1745. (Biology). View Reference
  5. Kojika S, Griffin JD. Notch receptors and hematopoiesis. Exp Hematol. 2001; 29(9):1041-1052. (Biology). View Reference
  6. Milner LA, Bigas A, Kopan R, Brashem-Stein C, Bernstein ID, Martin DI. Inhibition of granulocytic differentiation by mNotch1. Proc Natl Acad Sci U S A. 1996; 93(23):13014-13019. (Biology). View Reference
  7. Milner LA, Bigas A. Notch as a mediator of cell fate determination in hematopoiesis: evidence and speculation. Blood. 1999; 93(8):2431-2448. (Biology). View Reference
  8. 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 (8) View Less
552768 Rev. 2

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