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RB545 Mouse Anti-Human CD86 (B7-2)
RB545 Mouse Anti-Human CD86 (B7-2)

Multiparameter flow cytometric analysis of CD86 (B7-2) expression on Human peripheral blood leukocyte populations.  Human whole blood was first treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and then preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220). The leukocytes were then stained with either BD Horizon™ RB545 Mouse IgG1, κ Isotype Control (Cat. No. 569284; Left Plot) or BD Horizon™ RB545 Mouse Anti-Human CD86 (B7-2) antibody (Cat. No. 569734/569762; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD86 (B7-2) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leukocytes. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 software.

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¥67,760
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BD Horizon™
B7.2; B7-2; B-lymphocyte activation antigen B7-2; B70; CD28LG2; LAB72
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human ARH 77 Cell Line
Flow cytometry (Routinely Tested)
5 µl/test
V BP072, A109; VI BP95, CD86.9
942
AB_3685274
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation and Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

推奨アッセイ手順

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® 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 BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. For U.S. patents that may apply, see bd.com/patents.

データシート

569762 Rev. 1
抗体の詳細
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BU63

The BU63 monoclonal antibody specifically recognizes CD86 which is also known as B70, B7-2, B7.2, LAB72 or CD28LG2. CD86 is a ~70-80 kDa type I transmembrane glycoprotein that belongs to the Ig gene superfamily. CD86 is expressed on monocytes, dendritic cells, Langerhans cells, activated B cells, and memory B cells. Like CD80 (B7-1), CD86 serves as a ligand for CD28 or CD152 (CTLA-4) expressed by T cells and can induce costimulatory or coinhibitory signals, respectively. The BU63 antibody can reportedly block the binding of other human CD86-specific antibodies including the 2331 (FUN-1) (complete block) or IT2.2 (partial block) antibodies. These results suggest that the BU63 antibody may bind to a similar or identical CD86 epitope as the 2331 (FUN-1) antibody but to a different, although spatially-related epitope compared with the IT2.2 antibody.

569762 Rev. 1
フォーマットの詳細
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RB545
The BD Horizon RealBlue™ 545 (RB545) fluorochrome is part of the BD family of blue (488-nm) laser dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 496-nm and an emission maximum (Em Max) at 545-nm. Driven by BD innovation, RB545 can be used on spectral cytometers and is designed to be excited by the blue (488-nm) laser with minimal excitation by the 561-nm yellow-green laser. RB545 has minimal spillover into yellow-green detectors and a brightness level similar to FITC.  Given its unique emission max, on a spectral instrument RB545 can be used simultaneously with FITC and PE to provide an additional color off of the blue laser. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
altImg
RB545
Blue 488 nm
496 nm
545 nm
569762 Rev.1
引用&参考文献
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Development References (5)

  1. Azuma M, Ito D, Yagita H, et al. B70 antigen is a second ligand for CTLA-4 and CD28.. Nature. 1993; 366(6450):76-9. (Biology). View Reference
  2. Engel P, Gribben JG, Freeman GJ, et al. The B7-2 (B70) costimulatory molecule expressed by monocytes and activated B lymphocytes is the CD86 differentiation antigen. Blood. 1994; 84(5):1402-1407. (Clone-specific: Blocking, Flow cytometry). View Reference
  3. Engel P, Wagner N, Tedder TF. CD86 Workshop Report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:703-705.
  4. Hardie DL, Casamayor M, Johnson GD, et al. CD86 Workshop Panel report. In: 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.; 1997:201-204.
  5. Nozawa Y, Abe M, Wakasa H. Three mAb, FUN-1, FB1, and FB21, that recognize B-cell antigens in frozen or paraffin-embedded tissue sections. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:705-706.
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569762 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

 

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