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BUV395 Mouse Anti-Human CD146
BUV395 Mouse Anti-Human CD146

Flow cytometric analysis of CD146 expression on Human HeLa cells. Cells from the human HeLa (Cervical adenocarcinoma, ATCC CCL-2) cell line were stained with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; dashed line histogram) or BD Horizon BUV395 Mouse Anti-Human CD146 antibody (Cat. No. 564326/565981; solid line histogram). The fluorescence histogram showing CD146 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of CD146 expression on Human HeLa cells. Cells from the human HeLa (Cervical adenocarcinoma, ATCC CCL-2) cell line were stained with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; dashed line histogram) or BD Horizon BUV395 Mouse Anti-Human CD146 antibody (Cat. No. 564326/565981; solid line histogram). The fluorescence histogram showing CD146 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Horizon™
MCAM; MELCAM; MUC18; Gicerin; Melanoma cell adhesion molecule
Human (QC Testing)
Mouse IgG1, κ
Human Umbilical Vein Cells
Flow cytometry (Routinely Tested)
5 µl
VIII
4162
AB_2738748
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 BUV395 under optimum conditions, and unconjugated antibody and free BD Horizon BUV395 were removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565981 Rev. 1
Antibody Details
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P1H12

The P1H12 monoclonal antibody specifically binds to CD146. CD146 is a 118 kDa transmembrane glycoprotein also known as MCAM, MUC18, or Mel-CAM. CD146 is a member of the immunoglobulin superfamily strongly expressed by blood vessel endothelium and smooth muscle. CD146 is also expressed by angioblasts, mesenchymal stem cells, melanoma cells, intermediate trophoblasts and a subpopulation of activated T cells. The P1H12 monoclonal antibody has been reported to block endothelial cell adhesion that is observed very early in embryogenesis. It can be useful in the study of embryologic vasculogenesis. This antibody is suitable for immunohistochemical staining of acetone-fixed frozen tissue sections, immunoprecipitation and ELISA.

The antibody was conjugated to BD Horizon BUV395 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. With an Ex Max near 348 nm and an Em Max near 395 nm, BD Horizon BUV395 can be excited by the ultraviolet laser (355 nm) laser and detected with a 379/28 filter. This dye has been exclusively developed by BD Biosciences as an optimal dye for use on instruments equipped with the ultraviolet laser and has virtually no spillover into any other detector.

565981 Rev. 1
Format Details
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BUV395
The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV395
Ultraviolet 355 nm
348 nm
395 nm
565981 Rev.1
Citations & References
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Development References (7)

  1. Elshal MF, Khan SS, Takahashi Y, Solomon MA, McCoy JP, Jr. CD146 (Mel-CAM), an adhesion marker of endothelial cells, is a novel marker of lymphocyte subset activation in normal peripheral blood. Blood. 2005; 106(8):2923-2924. (Clone-specific: Flow cytometry). View Reference
  2. Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
  3. Sers C, Kirsch K, Rothbächer U, Riethmüller G, Johnson JP. Genomic organization of the melanoma-associated glycoprotein MUC18: implications for the evolution of the immunoglobulin domains. Proc Natl Acad Sci U S A. 1993; 90(18):8514-8518. (Biology). View Reference
  4. Shih IM, Elder DE, Hsu MY, Herlyn M. Regulation of Mel-CAM/MUC18 expression on melanocytes of different stages of tumor progression by normal keratinocytes. Am J Pathol. 1994; 145(4):837-845. (Biology). View Reference
  5. Shih IM. The role of CD146 (Mel-CAM) in biology and pathology. J Pathol. 1999; 189(1):4-11. (Biology). View Reference
  6. Solovey A, Lin Y, Browne P, Choong S, Wayner E, Hebbel R P. Circulating activated endothelial cells in sickle cell anemia. N Engl J Med. 1997; 337(22):1584-1590. (Immunogen: Cell separation, Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
  7. Solovey AN, Gui L, Chang L, Enenstein J, Browne PV, Hebbel RP. Identification and functional assessment of endothelial P1H12. J Lab Clin Med. 2001; 138(5):322-331. (Clone-specific: Activation, Functional assay, Inhibition, Stimulation). View Reference
View All (7) View Less
565981 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.