APC-H7 Rat IgG2a, κ Isotype Control
Clone R35-95 (RUO)
- Brand BD Pharmingen™
- Concentration 0.2 mg/ml
- Isotype Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
- Reactivity Mouse (QC Testing)
Isotype control, Flow cytometry (Routinely Tested)
- Immunogen Mouse Pooled Immunoglobulin
- Storage Buffer Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The R35-95 hybridoma was generated by hybridization of Y3 myeloma cells with spleen cells from LOU rats immunized with mouse immunoglobulins. The R35-95 hybridoma produces rat IgG2a, κ immunoglobulin that has no measurable reactivity with mouse immunoglobulins. The R35-95 immunoglobulin was selected as an isotype control following screening for low background binding on a variety of mouse and human tissues.
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.
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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- 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.
- 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.
An isotype control should be used at the same concentration as the antibody of interest (e.g., ≤ 1 µg/million cells for flow cytometry).