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BD Pharmingen™ APC-H7 Mouse Anti-Human CD69
Clone FN50 (also known as FN 50) (RUO)
Flow cytometric analysis for CD69 in stimulated Rhesus macaque (Left Panel) or human (Right Panel) peripheral blood mononuclear cells (PBMC). PBMC from Rhesus or human were stimulated for at least 4-6 hours with 20 ng/mL PMA (Sigma-Aldrich Cat. No. P-8139) and 250 ng/mL calcium ionophore A23187 (Sigma-Aldrich Cat. No. C-9275). Cells were then stained with either a APC-H7 Mouse IgG1, κ isotype control (Cat. No. 560167; dashed line histograms) or with the APC-H7 Mouse Anti-Human CD69 antibody (Cat. No. 560737; solid line histograms). Histograms were derived from gated events based on light scattering characteristics for lymphocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Flow cytometric analysis for CD69 in stimulated Rhesus macaque (Left Panel) or human (Right Panel) peripheral blood mononuclear cells (PBMC). PBMC from Rhesus or human were stimulated for at least 4-6 hours with 20 ng/mL PMA (Sigma-Aldrich Cat. No. P-8139) and 250 ng/mL calcium ionophore A23187 (Sigma-Aldrich Cat. No. C-9275). Cells were then stained with either a APC-H7 Mouse IgG1, κ isotype control (Cat. No. 560167; dashed line histograms) or with the APC-H7 Mouse Anti-Human CD69 antibody (Cat. No. 560737; solid line histograms). Histograms were derived from gated events based on light scattering characteristics for lymphocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Flow cytometric analysis for CD69 in stimulated Rhesus macaque (Left Panel) or human (Right Panel) peripheral blood mononuclear cells (PBMC). PBMC from Rhesus or human were stimulated for at least 4-6 hours with 20 ng/mL PMA (Sigma-Aldrich Cat. No. P-8139) and 250 ng/mL calcium ionophore A23187 (Sigma-Aldrich Cat. No. C-9275). Cells were then stained with either a APC-H7 Mouse IgG1, κ isotype control (Cat. No. 560167; dashed line histograms) or with the APC-H7 Mouse Anti-Human CD69 antibody (Cat. No. 560737; solid line histograms). Histograms were derived from gated events based on light scattering characteristics for lymphocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Flow cytometric analysis for CD69 in stimulated Rhesus macaque (Left Panel) or human (Right Panel) peripheral blood mononuclear cells (PBMC). PBMC from Rhesus or human were stimulated for at least 4-6 hours with 20 ng/mL PMA (Sigma-Aldrich Cat. No. P-8139) and 250 ng/mL calcium ionophore A23187 (Sigma-Aldrich Cat. No. C-9275). Cells were then stained with either a APC-H7 Mouse IgG1, κ isotype control (Cat. No. 560167; dashed line histograms) or with the APC-H7 Mouse Anti-Human CD69 antibody (Cat. No. 560737; solid line histograms). Histograms were derived from gated events based on light scattering characteristics for lymphocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Product Details
BD Pharmingen™
AIM; CLEC2C; EA1; GP32/28; Leu23; MLR-3; VEA; BL-AC/P26
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse IgG1, κ
Anti-µ stimulated human B lymphocytes
Flow cytometry (Routinely Tested)
5 µl
IV A91 (A091)
AB_1727508
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.
- 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.
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Cy is a trademark of GE Healthcare.
- 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 Solution 10X Concentrate IVD
Size
100 ML
Cat No.
349202
Lysing Buffer RUO
Size
100 ML
Cat No.
555899
APC-H7 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
560167
560737 Rev. 2
Antibody Details
FN50
The FN50 monoclonal antibody specifically binds to human CD69. CD69 is also known as activation-induced molecule (AIM), early activation antigen (EA-1), very early activation antigen (VEA), C-type lectin domain family 2 member C (CLEC2C), MLR-3, GP32/28 and Leu-23. CD69 is a transmembrane type II homodimer receptor. CD69 is comprised of disulfide-linked, differentially glycosylated core protein subunits that are approximately 28 and 34 kDa in size. Each subunit contains a C-type lectin domain. CD69 is expressed on activated T, B, and natural killer (NK) lymphocytes, thymocytes, neutrophils, eosinophils and platelets. In normal peripheral blood, a small and variable percentage of lymphocytes typically express detectable membrane CD69 antigen. Upon activation, CD69 antigen expression increases on lymphocytes. Peak CD69 expression generally occurs within 18 hours of activation, preceding the appearance of HLA-DR, IL-2Rα (CD25) and transferrin receptor (CD71). CD69 is highly expressed on the bright CD3+ subset of thymocytes. FN50 monoclonal antibody labels NK cells and most lymphocytes of the follicular mantle and perifollicular/interfollicular zone as well as germinal center T cells of lymph nodes and tonsils. Studies indicate that CD69 serves as a signaling receptor in the activation of a variety of cell types.
560737 Rev. 2
Format Details
APC-H7
The BD Pharmingen™ 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
659 nm
782 nm
560737 Rev.2
Citations & References
View product citations for antibody "560737" on CiteAb
Development References (2)
-
Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
-
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.
560737 Rev. 2
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.
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.