Flow cytometric analysis of CD80 expression on Raji cells. Human Raji cells were stained with either APC-H7 Mouse Anti-Human CD80 antibody (Cat. No. 561134; solid line histogram) or with a APC-H7 Mouse IgG1, κ Isotype Control (Cat. No. 560167; dashed line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed on a BD LSR™ II Flow Cytometry System.
Flow cytometric analysis of CD80 expression on Raji cells. Human Raji cells were stained with either APC-H7 Mouse Anti-Human CD80 antibody (Cat. No. 561134; solid line histogram) or with a APC-H7 Mouse IgG1, κ Isotype Control (Cat. No. 560167; dashed line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed on a BD LSR™ II Flow Cytometry System.
Product Details
BD Pharmingen™
B7.1; B7-1; Activation B7-1 antigen; B7; BB1; CD28LG; CD28LG1; LAB7; L307
Human (QC Testing)
Mouse C3H, also known as C3H/He, C3H/Bi IgG1, κ
Human B7-transfected L cells
Flow cytometry (Routinely Tested)
5 µl
V B7.5
941
AB_10565974
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
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- 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.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561134 Rev. 2
Antibody Details
L307.4
The L307 monoclonal antibody specifically binds to B7/BB1, a 60 kDa transmembrane glycoprotein that was clustered as CD80 in the Fifth International Workshop on Human Leukocyte Differentiation Antigens. CD80, a member of the Ig supergene family, is expressed on activated B cells, T cells, macrophages, and dendritic cells. It is the ligand for two molecules expressed on T cells, CD28 and CD152 (CTLA-4). CD80 is also expressed on activated CD4-positive and CD8-positive T cells, appearing late after activation suggesting that activated T cells may be capable of autocrine costimulation via the CD28 activation pathway. The binding of CD28 by anti-CD28 or by CD80 results in T-cell activation and a signal for IL-2 production.
561134 Rev. 2
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
561134 Rev.2
Citations & References
View product citations for antibody "561134" on CiteAb
Development References (6)
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Azuma M, Yssel H, Phillips JH, Spits H, Lanier LL. Functional expression of B7/BB1 on activated T lymphocytes. J Exp Med. 1993; 177(3):845-850. (Biology). View Reference
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Behrens L, Kerschensteiner M, Misgeld T, Goebels N, Wekerle H, Hohlfeld R. Human muscle cells express a functional costimulatory molecule distinct from B7.1 (CD80) and B7.2 (CD86) in vitro and in inflammatory lesions. J Immunol. 1998; 161(11):5943-5951. (Biology). View Reference
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Freeman GJ, Cardoso AA, Boussiotis VA, et al. The BB1 monoclonal antibody recognizes both cell surface CD74 (MHC class II-associated invariant chain) as well as B7-1 (CD80), resolving the question regarding a third CD28/CTLA-4 counterreceptor. J Immunol. 1998; 161(6):2708-2715. (Biology). View Reference
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Koulova L, Clark EA, Shu G, Dupont B. The CD28 ligand B7/BB1 provides costimulatory signal for alloactivation of CD4+ T cells. J Exp Med. 1991; 173(3):759-762. (Biology). View Reference
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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.
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Schwartz RH. Costimulation of T lymphocytes: the role of CD28, CTLA-4, and B7/BB1 in interleukin-2 production and immunotherapy. Cell. 1992; 71(7):1065-1068. (Biology). View Reference
561134 Rev. 2
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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.