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Alexa Fluor® 647 Mouse Anti-Human VHL
Alexa Fluor® 647 Mouse Anti-Human VHL
Flow cytometric analysis of VHL expression in VHL-transfected 293F cells. Untransfected (dashed line histogram) and human VHL-transfected (solid line histogram) 293F cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then washed and stained with Alexa Fluor® 647 Mouse Anti-Human VHL antibody (Cat. No. 564184) using BD Biosciences Protocol for Intracellular Staining. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
Flow cytometric analysis of VHL expression in VHL-transfected 293F cells. Untransfected (dashed line histogram) and human VHL-transfected (solid line histogram) 293F cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then washed and stained with Alexa Fluor® 647 Mouse Anti-Human VHL antibody (Cat. No. 564184) using BD Biosciences Protocol for Intracellular Staining. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
Product Details
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BD Pharmingen™
VHL1; pVHL; Elongin binding protein; Protein G7; HRCA1; RCA1
Human (QC Testing)
Mouse IgG2b, κ
Human VHL Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2738652
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  3. 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.
  4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  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.
564184 Rev. 1
Antibody Details
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S2-647

The S2-647 monoclonal antibody recognizes the Von Hippel-Lindau (VHL) protein. The VHL protein is expressed within the cytosol and nuclei of cells from a variety of tissues. It serves as a tumor suppressor and transcriptional repressor that is highly conserved from Drosophila to mammalian species.  VHL binds to Elongin B and C subunits and other proteins as part of a E3 ubiquitin ligase complex that targets a variety of proteins, including HIF-1 alpha and Beta-2 adrenergic receptors, for proteasomal degradation. Aberrant VHL expression is associated with a number of diseases including Von Hippel-Lindau disease, a familial cancer syndrome, that predisposes individuals to hemangioblastomas of the central nervous system and retina, pheochromocytoma and renal cell carcinoma.  Three distinct VHL isoforms ranging in molecular weight from approximately 19-30 kDa have been described.

  

564184 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
564184 Rev.1
Citations & References
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Development References (8)

  1. Baba M, Hirai S, Kawakam S, et al. Tumor suppressor protein VHL is induced at high cell density and mediates contact inhibition of cell growth. Oncogene. 2001; 20(22):2727-2736. (Biology). View Reference
  2. Chen F, Kishida T, Duh FM, et al. Suppression of growth of renal carcinoma cells by the von Hippel-Lindau tumor suppressor gene. Cancer Res. 1995; 55(21):4804-4807. (Biology). View Reference
  3. Hsu T, Adereth Y, Kose N, Dammai V. Endocytic function of von Hippel-Lindau tumor suppressor protein regulates surface localization of fibroblast growth factor receptor 1 and cell motility. J Biol Chem. 2006; 28(281):12069-12080. (Biology). View Reference
  4. Iliopoulos O, Kibel A, Gray S, Kaelin WG Jr. Tumour suppression by the human von Hippel-Lindau gene product.. Nat Med. 1995; 1(8):822-826. (Biology). View Reference
  5. Iliopoulos O, Ohh M, Kaelin WG Jr.. pVHL19 is a biologically active product of the von Hippel-Lindau gene arising from internal translation initiation. Proc Natl Acad Sci U S A. 1998; 95(20):11661-11666. (Biology). View Reference
  6. In: Hesketh R. The Oncogene Handbook. New York: Academic Press; 1994:559.
  7. Kibel A, Iliopoulos O, DeCaprio JA, Kaelin WG Jr. Binding of the von Hippel-Lindau tumor suppressor protein to Elongin B and C. Science. 1995; 269(5229):1444-1446. (Biology). View Reference
  8. Qi H, Gervais ML, Li W, DeCaprio JA, Challis JR, Ohh M. Molecular cloning and characterization of the von Hippel-Lindau-like protein. Mol Cancer Res. 2004; 2(1):43-52. (Biology). View Reference
View All (8) View Less
564184 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.