BD Pharmingen™ PE Rat IgG2a κ Isotype Control

I/C Flow Cytometry: The PE-conjugated R35-95 immunoglobulins (Cat. No. 559317) is a suitable rat IgG2a κ isotype controls for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse or human cells for flow cytometric analysis.

Important Note: This pre-titered antibody solution does not contain a cell permeabilization agent. It is necessary to include a cell permeabilization agent when using the pre-titered antibody solution to stain fixed and permeabilized cells. BD Perm/Wash™ Buffer (Cat. No 554723) contains the permeabilization agent saponin and is useful for this purpose as described in the Protocol section below.

Protocol

1. Resuspend 1 x 10^6 fixed and permeabilized cells in 20 µl of the pre-titered antibody solution and 30 µl of 1X Perm/Wash Buffer (Cat. No 554723).

2. Incubate the cell suspension for 15 minutes (at RT or 4°C).

3. Wash twice in 100 µl of 1X Perm/Wash Buffer (Cat. No 554723).

1. 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).
2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
3. An isotype control should be used at the same concentration as the antibody of interest.
4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
6. 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.
7. Source of all serum proteins is from USDA inspected abattoirs located in the United States.