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BV421 Mouse Anti-Human CD243
BV421 Mouse Anti-Human CD243
Multicolor flow cytometric analysis of CD243 expression on human neuroblastoma cells. Cells from the human SH-SY5Y (Neuroblastoma, ATCC  CRL-2266) cell line were stained with either BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; dashed line histogram) or BD Horizon BV421 Mouse Anti-Human CD243 antibody (Cat. No. 566015; solid line histogram). The fluorescence histogram showing CD243 expression (or Ig Isotype control staining), were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Multicolor flow cytometric analysis of CD243 expression on human neuroblastoma cells. Cells from the human SH-SY5Y (Neuroblastoma, ATCC  CRL-2266) cell line were stained with either BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; dashed line histogram) or BD Horizon BV421 Mouse Anti-Human CD243 antibody (Cat. No. 566015; solid line histogram). The fluorescence histogram showing CD243 expression (or Ig Isotype control staining), were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Product Details
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BD Horizon™
ABCB1; ABC20; CLCS; GP170; MDR1; P-GP; PGY1
Human (QC Testing)
Mouse BALB/c IgG2a, κ
Human MDR1 Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
VII 70364
AB_2739460
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.

Recommended Assay Procedures

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).

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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  7. An isotype control should be used at the same concentration as the antibody of interest.
566015 Rev. 1
Antibody Details
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UIC2

The UIC2 monoclonal antibody specifically binds to an extracellular epitope of CD243, which is also known as ATP-binding cassette subfamily B member 1 (ABCB1), Multidrug resistance protein 1 (MDR1), or P-glycoprotein 1. CD243 is a transmembrane glycoprotein that spans the membrane 12 times. CD243 acts as an ATP-dependent efflux pump for a large variety of lipophilic molecules and drugs. This efflux activity has been suggested to lead to resistance to the drugs used in chemotherapy. CD243 is expressed by epithelial and endothelial cells, and at low levels by T cells, B cells, NK cells, and hematopoietic stem cells. It may be expressed at high levels by multidrug resistant (MDR) tumor cells. The UIC2 antibody reportedly inhibited the efflux of CD243 substrates from MDR cells and increased the cytotoxicity of certain CD243-transported drugs. The UIC2 antibody reportedly does not crossreact with mouse CD243.

 

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.

566015 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
566015 Rev.1
Citations & References
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Development References (5)

  1. Mechetner EB1, Roninson IB. Efficient inhibition of P-glycoprotein-mediated multidrug resistance with a monoclonal antibody.. Proc Natl Acad Sci U S A. 1992; 89(13):5824-5828. (Immunogen: Blocking, Flow cytometry, Functional assay, Immunoprecipitation, Inhibition, Radioimmunoassay). View Reference
  2. Nagy H, Goda K, Arceci R, Cianfriglia M, Mechetner E, Szabó G. P-Glycoprotein conformational changes detected by antibody competition.. Eur J Biochem. 2001; 268(8):2416-20. (Clone-specific: Flow cytometry). View Reference
  3. Ritchie TK, Kwon H, Atkins WM. Conformational analysis of human ATP-binding cassette transporter ABCB1 in lipid nanodiscs and inhibition by the antibodies MRK16 and UIC2.. J Biol Chem. 2011; 286(45):39489-96. (Clone-specific: Functional assay, Inhibition). View Reference
  4. Szalóki G, Krasznai ZT, Tóth Á, et al. The strong in vivo anti-tumor effect of the UIC2 monoclonal antibody is the combined result of Pgp inhibition and antibody dependent cell-mediated cytotoxicity.. PLoS ONE. 2014; 9(9):e107875. (Clone-specific: Cytotoxicity, Functional assay). View Reference
  5. Zhou Y, Gottesman MM, Pastan I. The extracellular loop between TM5 and TM6 of P-glycoprotein is required for reactivity with monoclonal antibody UIC2.. Arch Biochem Biophys. 1999; 367(1):74-80. (Clone-specific: Functional assay, Inhibition). View Reference
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566015 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.