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BB515 Mouse Anti-Human CD70
BB515 Mouse Anti-Human CD70
Flow cytometric analysis of CD70 expression on human U266 myeloma cells - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Cells from the human U266 (Myeloma, ATCC TIB-196) cell line were stained with BD Horizon™ BB515 Mouse IgG3, κ Isotype Control (Cat. No. 565114; dashed line histogram) or BD Horizon™ BB515 Mouse Anti-Human CD70 antibody (Cat. No. 565156; bold solid line histogram). Alternatively, cells were stained with FITC Anti-Human CD70 antibody (Cat. No. 555834; thin solid line histogram).        Overlaid histograms are shown to facilitate staining comparisons between: BB515 Anti-CD70 antibody versus its Ig Isotype Control (Left Panel), and BB515 Anti-CD70 antibody versus FITC Anti-CD70 antibody (Right Panel). The fluorescence histograms showing CD70 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable U266 cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Flow cytometric analysis of CD70 expression on human U266 myeloma cells - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Cells from the human U266 (Myeloma, ATCC TIB-196) cell line were stained with BD Horizon™ BB515 Mouse IgG3, κ Isotype Control (Cat. No. 565114; dashed line histogram) or BD Horizon™ BB515 Mouse Anti-Human CD70 antibody (Cat. No. 565156; bold solid line histogram). Alternatively, cells were stained with FITC Anti-Human CD70 antibody (Cat. No. 555834; thin solid line histogram).        Overlaid histograms are shown to facilitate staining comparisons between: BB515 Anti-CD70 antibody versus its Ig Isotype Control (Left Panel), and BB515 Anti-CD70 antibody versus FITC Anti-CD70 antibody (Right Panel). The fluorescence histograms showing CD70 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable U266 cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Horizon™
CDw70; CD27 ligand; CD27-L; CD27L; CD27LG; Ki-24 antigen; TNFSF7
Human (QC Testing)
Mouse IgG3, κ
Human L428 Cell Line
Flow cytometry (Routinely Tested)
5 µl
III 166; IV A109
AB_2739083
Aqueous buffered solution containing ≤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™ BB515 under optimum conditions and unconjugated antibody was 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).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. 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.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565156 Rev. 3
Antibody Details
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Ki-24

The Ki-24 monoclonal antibody specifically binds to human CD70. CD70 is a type II transmembrane glycoprotein and member of the TNF Superfamily. CD70 is also known as Tumor necrosis factor ligand superfamily member 7 (TNFSF7), CD27 ligand (CD27-L, CD27L, CD27LG), and KI-24 antigen. The CD70 antigen immunoprecipitates as five bands (50, 70, 90, 100 and 160 kDa) under non-reducing conditions. CD70 is strongly expressed on Reed-Sternberg cells, some activated T or B cells and Epstein Barr Virus (EBV)-positive lymphoblastoid cell lines. CD70 plays roles in the activation, proliferation and differentiation of B cells and T cells including the enhanced production of cytotoxic T cells.

The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

565156 Rev. 3
Format Details
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BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BB515
Blue 488 nm
490 nm
515 nm
565156 Rev.3
Citations & References
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Development References (8)

  1. Bowman MR, Crimmins MA, Yetz-Aldape J, Kriz R, Kelleher K, Herrmann S. The cloning of CD70 and its identification as the ligand for CD27. J Immunol. 1994; 152(4):1756-1761. (Biology). View Reference
  2. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  3. 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.
  4. Stein H, Ferszt A, Dallenbach F, et al. CDw70 mAb A109 (Ki-24): expression by reactive and neoplastic lymphoid cells. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:449-451.
  5. Stein H, Gerdes J, Lemke H, Mason DY. Evidence of Sternberg-Reed cells being derived from activated lymphocytes. Haematol Blood Transfus. 1985; 29:441-444. (Clone-specific: Immunocytochemistry (cytospins)). View Reference
  6. Stein H, Gerdes J, Schwab U, et al. Evidence for the detection of the normal counterpart of Hodgkin and Sternberg-Reed cells.. Hematol Oncol. 1(1):21-9. (Clone-specific). View Reference
  7. Stein H, Gerdes J, Schwarting R, Froese P, Lemke H. Three new lymphoid activation antigens. In: McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:574.
  8. Stein H, Schwarting R, Niedobitek G, Dallenbach F. Cluster report: CDw70. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:446-449.
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565156 Rev. 3

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