APC-H7 Mouse Anti-Human CD69
Clone FN50 (also known as FN 50) (RUO)
- Brand BD Pharmingen™
- Alternative Name AIM; CLEC2C; EA1; GP32/28; Leu23; MLR-3; VEA; BL-AC/P26
- Vol. Per Test 5 µl
- Isotype Mouse IgG1, κ
- Reactivity Human (QC Testing) Rhesus, Cynomolgus, Baboon (Tested in Development)
Flow cytometry (Routinely Tested)
- Immunogen Anti-µ stimulated human B lymphocytes
- Workshop No. IV A91 (A091)
- Entrez Gene ID 969
- Storage Buffer Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
APC-H7 is an APC-cyanine tandem fluorochrome, which uses an analog of Cy7 and has similar spectral properties to APC-Cy7. APC-H7 conjugates provide greater stability in light and paraformaldehyde fixatives and have less spillover into the APC channel than APC-Cy7 conjugates. APC-H7 conjugates are typically 75% as bright as equivalent APC-Cy7 conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, APC-Cy7 and APC-H7 cannot be used simultaneously.
Suggested Companion Products
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.
- 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 testing during development may have been performed with a different format of the same clone. Selected applications have been tested for cross-reactivity.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.