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BV421 Mouse Anti-Human Notch4
BV421 Mouse Anti-Human Notch4

Flow Cytometric Analysis of Notch4 expression on a human lymphoblastic leukemia cell line. Cells from the human MOLT-3 (T lymphoblastic leukemia, ATCC CRL-1552) cell line were harvested and stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human Notch4 antibody (Cat. No. 563905; solid line histogram) at matched concentrations. The fluorescence histograms were derived from gated events with the forward and side light scattering characteristics of viable MOLT-3 cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Flow Cytometric Analysis of Notch4 expression on a human lymphoblastic leukemia cell line. Cells from the human MOLT-3 (T lymphoblastic leukemia, ATCC CRL-1552) cell line were harvested and stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human Notch4 antibody (Cat. No. 563905; solid line histogram) at matched concentrations. The fluorescence histograms were derived from gated events with the forward and side light scattering characteristics of viable MOLT-3 cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Horizon™
Notch-4; Neurogenic locus notch homolog protein 4; hNotch4; NOTC4; INT3
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human Notch4 Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
AB_2738478
Aqueous buffered solution containing BSA 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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. Brilliant Violet™ 421 is a trademark of Sirigen.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563905 Rev. 1
Antibody Details
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MHN4-2

The MHN4-2 monoclonal antibody specifically binds to an extracellular domain of Neurogenic locus notch homolog protein 4, also known as, Notch4 (or Notch 4 and Notch-4). Notch4 is a type 1 transmembrane glycoprotein that belongs to the Notch Receptor Family comprised of Notch1 through Notch4. Notch4 serves as a receptor for several ligands including Delta1, Jagged1, and Jagged2 that can be expressed on the same or different cells. Notch4 plays important roles in embryonic and postnatal development and cell fate determination by regulating cellular differentiation, proliferation and apoptosis. After ligand binding, the Notch4 receptor is cleaved in its extracellular and transmembrane domains. The cleaved notch intracellular domain (NICD) dissociates from the membrane and translocates to the nucleus where it can act as a transcriptional activator of downstream target genes involved with cellular differentiation and other responses. Notch4 is expressed at high levels in the heart and by endothelial cells and by certain CD34-positive and CD34-negative human bone marrow cell subsets. It is over-expressed by cells in certain breast, colon, and lung cancers.

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.

563905 Rev. 1
Format Details
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BV421
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.
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BV421
Violet 405 nm
407 nm
423 nm
563905 Rev.1
Citations & References
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Development References (8)

  1. Harrison H, Farnie G, Howell SJ, et al. Regulation of breast cancer stem cell activity by signaling through the Notch4 receptor. Cancer Res. 2010; 70(2):709-718. (Biology). View Reference
  2. Karanu FN, Yuefei L, Gallacher L, Sakano S, Bhatia M. Differential response of primitive human CD34- and CD34+ hematopoietic cells to the Notch ligand Jagged-1. Leukemia. 2003; 17(7):1366-1374. (Biology). View Reference
  3. Li L, Huang GM, Banta AB, et al. Cloning, characterization, and the complete 56.8-kilobase DNA sequence of the human NOTCH4 gene. Genomics. 1998; 51:45-58. (Biology). View Reference
  4. Sekine C, Koyanagi A, Koyama N, Hozumi K, Chiba S, Yagita H. Differential regulation of osteoclastogenesis by Notch2/Delta-like 1 and Notch1/Jagged1 axes. Arthritis Res Ther. 2012; 14(2):R45-R58. (Immunogen: Flow cytometry, Functional assay). View Reference
  5. Sugaya K, Sasanuma S, Nohata J, et al. Gene organization of human NOTCH4 and (CTG)n polymorphism in this human counterpart gene of mouse proto-oncogene Int3. Gene. 1997; 189(2):235-244. (Biology). View Reference
  6. Sun Y, Lowther W, Kato K, et al. Notch4 intracellular domain binding to Smad3 and inhibition of the TGF-beta signaling. Oncogene. 2005; 24(34):5365-5374. (Biology). View Reference
  7. Vercauteren SM, Sutherland HJ. Constitutively active Notch4 promotes early human hematopoietic progenitor cell maintenance while inhibiting differentiation and causes lymphoid abnormalities in vivo. Blood. 2004; 104(8):2315-2322. (Biology). View Reference
  8. Wu J, Iwata F, Grass JA, et al. Molecular determinants of NOTCH4 transcription in vascular endothelium. Mol Cell Biol. 2005; 25(4):1458-1474. (Biology). View Reference
View All (8) View Less
563905 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.