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Alexa Fluor® 647 Mouse Anti-Human Notch4
Alexa Fluor® 647 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 Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human Notch4 antibody (Cat. No. 563904; solid line histogram) at matched concentrations. The fluorescence histograms were derived from gated events based on 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 Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human Notch4 antibody (Cat. No. 563904; solid line histogram) at matched concentrations. The fluorescence histograms were derived from gated events based on 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 Pharmingen™
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_2738477
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

  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. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. 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.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  7. 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.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563904 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.

563904 Rev. 1
Format Details
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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
563904 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
563904 Rev. 1

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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.