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BD Pharmingen™ Purified Mouse Anti-Rat CD32
Clone D34-485 (RUO)
Two-color analysis of the expression of Cd32 on rat splenocytes. Lewis rat splenocytes were stained with purified D34-485 mAb, followed by FITC-conjugated polyclonal goat anti-mouse Ig (Cat. No. 554001), then PE-conjugated anti-rat IgM G35-238 (Cat. No. 553888). Double-positive cells are B lymphocytes (IgM+ cells), which express CD32 on the cell surface. Flow cytometry was performed on a BD FACScan™ flow cytometry system.
Blocking of Fc-mediated binding to FcγII receptors (CD32) on rat splenocytes. Lewis rat splenocytes were pre-incubated with purified isotype control mAb A112-2 (Cat. No. 553487, left panel) or Rat BD Fc Block™ purified anti-rat CD32 mAb D34-485 (right panel). Two-color staining was performed with FITC-conjugated anti-rat NKR-P1A mAb 10/78 (Cat. No. 555008) and PE-conjugated anti-rat CD45RA mAb OX-33 (Cat. No. 551402/554884). Note how the dim staining of B lymphocytes (OX-33+ cells) by anti-CD161a (NKR-P1A) (left panel) is reduced when Rat BD Fc Block™ is used before staining (right panel). Flow cytometry was performed on a BD FACScan™ flow cytometry system.
Two-color analysis of the expression of Cd32 on rat splenocytes. Lewis rat splenocytes were stained with purified D34-485 mAb, followed by FITC-conjugated polyclonal goat anti-mouse Ig (Cat. No. 554001), then PE-conjugated anti-rat IgM G35-238 (Cat. No. 553888). Double-positive cells are B lymphocytes (IgM+ cells), which express CD32 on the cell surface. Flow cytometry was performed on a BD FACScan™ flow cytometry system.
Blocking of Fc-mediated binding to FcγII receptors (CD32) on rat splenocytes. Lewis rat splenocytes were pre-incubated with purified isotype control mAb A112-2 (Cat. No. 553487, left panel) or Rat BD Fc Block™ purified anti-rat CD32 mAb D34-485 (right panel). Two-color staining was performed with FITC-conjugated anti-rat NKR-P1A mAb 10/78 (Cat. No. 555008) and PE-conjugated anti-rat CD45RA mAb OX-33 (Cat. No. 551402/554884). Note how the dim staining of B lymphocytes (OX-33+ cells) by anti-CD161a (NKR-P1A) (left panel) is reduced when Rat BD Fc Block™ is used before staining (right panel). Flow cytometry was performed on a BD FACScan™ flow cytometry system.
Product Details
BD Pharmingen™
Fcgr2b; FcRII; Fc-gamma RII; FcγRII; FcγII receptor
Rat (QC Testing)
Mouse BALB/c IgG1, κ
Recombinant Rat CD32 Protein
Flow cytometry (Routinely Tested), Blocking, Immunohistochemistry-frozen, Immunohistochemistry-zinc-fixed, Western blot (Reported), Immunohistochemistry-paraffin (Not Recommended)
0.5 mg/ml
AB_393567
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
Recommended Assay Procedures
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)
Product Notices
- 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.
- 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.
- 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.
Companion Products
Purified Mouse IgG1, κ Isotype Control RUO
Size
0.5 mg
Cat No.
557273
FITC Goat Anti-Mouse Ig RUO
Size
0.5 mg
Cat No.
554001
PE Mouse Anti-Rat IgM RUO
Size
0.2 mg
Cat No.
553888
FITC Mouse Anti-Rat CD161a RUO
Size
0.5 mg
Cat No.
555008
FITC Goat Anti-Mouse IgG/IgM RUO
Size
0.5 mg
Cat No.
555988
PE Mouse Anti-Rat CD45RA RUO
Size
0.1 mg
Cat No.
551402
550270 Rev. 5
Antibody Details
D34-485
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.
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.
550270 Rev. 5
Format Details
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
550270 Rev.5
Citations & References
View product citations for antibody "550270" on CiteAb
No Citations Are Available for this Product
550270 Rev. 5
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.