BD Pharmingen™ Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™)
Clone 2.4G2
Preparation And Storage
Recommended Assay Procedures
To specifically stain cells bearing FcγII and FcγIII 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 or Fc receptor-mediated binding by PE-CY5 tandem dye conjugates to FcγII and FcγIII receptor-bearing mouse cells for flow cytometric analysis:
1. Preincubate cell suspension with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (eg, ≤ 1 μg/million cells in 100 μl) at 4˚C for 5 minutes.
2. Add antibody of interest directly to preincubated cells in the presence of Mouse BD Fc Block™ (ie, Mouse BD Fc Block™ need not be washed off before staining cells).
3. If anti-Ig second-step is necessary, a reagent must be chosen which will not bind to Mouse BD Fc Block™ (eg, rat IgG2b, κ).
For additional information on using Mouse BD Fc Block™, refer to our website protocol at http://www.bdbiosciences.com/pharmingen/protocols/Immunophenotyping.shtml
The 2.4G2 antibody specifically recognizes a common nonpolymorphic epitope on the extracellular domains of the mouse FcγIII (CD16) and FcγII (CD32) Receptors. It has also been reported to bind the FcγI receptor (CD64) via its Fc domain. 2.4G2 mAb blocks non-antigen-specific binding of immunoglobulins to the FcγIII and FcγII, and possibly FcγI, Receptors in vitro and in vivo. CD16 and/or CD32 are expressed on natural killer cells, monocytes, macrophages, dendritic cells (at low levels), Kupffer cells, granulocytes, mast cells, B lymphocytes, immature thymocytes, and some activated mature T lymphocytes.
This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.