BUF041B is a rat antibody that recognises mouse CD16 and CD32, cell surface proteins also known as FcRgammaIII and FcRgammaII, respectively.
The function of these proteins is to bind IgG molecules via their Fc regions as part of the adaptive immune response. CD16 and/or CD32 are expressed by a wide variety of cells, including monocytes, macrophages, B lymphocytes, granulocytes, NK cells, dendritic cells and some activated T lymphocytes.
The expression of CD16/CD32 antigens can lead to non-specific binding of test monoclonal antibodies in staining procedures, resulting in “high background” staining on a wide range of cells. This non-specific binding may be blocked by pre-incubation of target cells with BUF041B, resulting in clearer staining.
For direct analysis of CD16/32 expression this antibody is also available conjugated directly to FITC (MCA2305F), AlexaFluor488 (MCA2305A488), AlexaFluor647 (MCA2305A647), Biotin (MCA2305B) and RPE (MCA2305PE).
Application
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
Flow Cytometry
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Storage
Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
18 months from date of despatch.
Instructions For Use
In order to reduce Fc-receptor mediated binding of test antibodies the following procedure is recommended:-
1) Incubate the cell suspension (1 x 106cells in 100ul) with 1ug of BUF041B (1ul of undiluted reagent, or 10ul of a 1/10 dilution) for 5-10 minutes.
2) Add test antibody according to manufacturers instructions – Do not wash BUF041B off the cells. BUF041B is suitable for use in conjunction with test antibodies from any manufacturer or with in-house antibodies.*
3) Proceed with staining as usual.
* Care is needed in the design of experiments utilizing unconjugated anti-mouse antibodies, to ensure that the secondary antibody being used does not cross-react with SeroBlock FcR. AbD Serotec supply a range of isotype specific anti-mouse immunoglobulin antibodies that may be useful for this purpose.
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