Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™)
Clone 2.4G2 (RUO)
- Brand BD Pharmingen™
- Alternative Name FcγRIII/FcγRII; Fcgr3/Fcgr2
- Concentration 0.5 mg/ml
- Isotype Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
- Reactivity Mouse (QC Testing)
Flow cytometry, Blocking (Routinely Tested)
Immunohistochemistry-frozen (Tested During Development)
- Immunogen Mouse BALB/c Macrophage J774
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Store undiluted at 4°C.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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