Description
The D34-485 monoclonal antibody specifically recognizes CD32, the FcγII receptor. Rat CD32 is expressed on B lymphocytes, myeloid cells, and some lymphocytes in the thymic medulla. D34-485 antibody blocks binding of aggregated immunoglobulins to the FcγII receptors in vitro.
To specifically stain cells bearing FcγII receptors for flow cytometric analysis: Incubate cell suspension with this antibody (≤ 1 µg/million cells) followed by an appropriate fluorochrome-conjugated second-step reagent. To reduce Fc receptor-mediated binding by antibodies of interest to FcγII receptor-bearing rat cells for flow cytometric analysis: a. Preincubate cell suspension with Rat BD Fc Block™ (e.g., ≤ 1 µg/million cells in 100 µl) at 4°C for 5 minutes. b. Add antibody of interest directly to preincubated cells in the presence of Rat BD Fc Block™ (i.e., Rat BD Fc Block™ need not be washed off before staining cells) c. If anti-Ig second-step is necessary, a reagent must be chosen which will not bind to Rat BD Fc Block™ (e.g., mouse IgG1, κ)
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. 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.
4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
