Flow cytometric analysis of CD27 expression on human peripheral blood lymphocytes. Whole blood was stained with either APC-H7 Mouse IgG1, κ Isotype Control (Cat. No. 560167; dashed line histogram) or APC-H7 Mouse Anti-Human CD27 (Cat. No. 560222/560223; solid line histogram). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Fluorescence histograms depicting CD27 (or Ig isotype) expression were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes. Flow cytometry was performed on a BD™ LSR II system.
Flow cytometric analysis of CD27 expression on human peripheral blood lymphocytes. Whole blood was stained with either APC-H7 Mouse IgG1, κ Isotype Control (Cat. No. 560167; dashed line histogram) or APC-H7 Mouse Anti-Human CD27 (Cat. No. 560222/560223; solid line histogram). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Fluorescence histograms depicting CD27 (or Ig isotype) expression were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes. Flow cytometry was performed on a BD™ LSR II system.
Product Details
BD Pharmingen™
TNFRSF7; TNF receptor superfamily, member 7; T14; Tp55; S152
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse BALB/c IgG1, κ
Human T-CLL cells
Flow cytometry (Routinely Tested)
5 µl
IV T187; V 5T CD27.03
939
AB_1645474
Aqueous buffered solution containing BSA, protein stabilizer, 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 APC-H7 under optimum conditions, and unconjugated antibody and APC-H7 were removed.
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- BD APC-H7 is a tandem conjugate and an analog of APC-Cy7 with the same spectral properties. It has decreased intensity but it is engineered for greater stability and less spillover in the APC channel and consequently offers better performance than APC-Cy7. It has an absorption maximum of approximately 650 nm. When excited by light from a red laser, the APC fluorochrome can transfer energy to the cyanine dye, which then emits at a longer wavelength. The resulting fluorescent emission maximum is approximately 767 nm. BD recommends that a 750-nm longpass filter be used along with a red-sensitive detector such as the Hamamatsu R3896 PMT. As with APC-Cy7 special filters are required when using APC-H7 in conjunction with APC. Note: Although our APC-H7 products demonstrate higher lot-to lot consistency than other APC tandem conjugate products, and every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-H7 conjugate.
- Although BD APC-H7 is engineered to minimize spillover to the APC channel and is more stable and less affected by light, temperature, and formaldehyde-based fixatives, compared to other APC-cyanine tandem dyes, it is still good practice to minimize as much as possible, any light, temperature and fixative exposure when working with all fluorescent conjugates.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of GE Healthcare.
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
APC-H7 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
560167
560222 Rev. 1
Antibody Details
M-T271
The M-T271 monoclonal antibody specifically binds to CD27. CD27 presents as a type I transmembrane, disulphide-linked 110 kDa homodimer comprised of two polypeptide chains. The CD27 molecule is a lymphocyte-specific member of the TNF/NGF-R family, and is expressed on a subset of human thymocytes and on the majority of mature T lymphocytes, activated B cells and NK cells. CD27 is highly induced on T cells after TCR stimulation. CD27 binds to CD70 (also known as, CD27 ligand or CD27L) and may be involved in cellular interaction of T and B lymphocytes.
560222 Rev. 1
Format Details
APC-H7
The BD Horizon™ APC-H7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 659 nm and an acceptor dye, H7, with an emission maximum (Em Max) at 782 nm. APC-H7, driven by BD innovation, is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC-H7
Red 627-640 nm
659 nm
782 nm
560222 Rev.1
Citations & References
View product citations for antibody "560222" on CiteAb
Development References (4)
-
Bigler RD, Bushkin Y, Chiorazzi N. S152 (CD27). A modulating disulfide-linked T cell activation antigen. J Immunol. 1988; 141(1):21-28. (Biology). View Reference
-
Bigler RD, Donat TL, Boselli CM. Definition of three epitopes of the CD27 molecule [P 120->55] present on activated normal lymphocytes. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:351-352.
-
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.
-
Watts TH. TNF/TNFR family members in costimulation of T cell responses. Annu Rev Immunol. 2005; 23:23-68. (Biology). View Reference
560222 Rev. 1
Please refer to Support Documents for Quality Certificates
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.