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Flow cytometric analysis of CD22.2 expression by mouse splenocytes using PE Rat Anti-Mouse IgG1 as a second step. BALB/c mouse splenic leucocytes were stained with either Purified Mouse IgG1, κ Anti-Mouse CD22.2 antibody (clone Cy34.1; solid line histogram) or with no antibody (for background staining control; dashed line histogram). After washing, the cells were stained with PE Rat Anti-Mouse IgG1 antibody (Cat. No. 550083) as the second step at 1 µg/test. The fluorescence histogram showing CD22.2 expression (or control staining) as detected by PE Rat Anti-Mouse IgG1 antibody was derived from gated events with the forward and side light-scatter characteristics of intact splenocytes. Flow cytometry and analysis was performed using a BD™ LSR II Flow Cytometer System and FlowJo™ software.
BD Pharmingen™ PE Rat Anti-Mouse IgG1
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
For flow cytometric detection of intracytoplasmic IgG1, we recommend FITC-conjugated mAb A85-1 (Cat. No. 553443).
BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
The A85-1 antibody reacts specifically with mouse IgG1 of Igh-Ca and Igh-Cb haplotypes. It does not react with other Ig isotypes. Detection of surface immunoglobulin on B lymphoma cells has been demonstrated with the A85-1 monoclonal antibody. A suspension of pooled mouse IgG1 was used as the source of immunogen.
Development References (3)
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Bhattacharya D, Lee DU, Sha WC. Regulation of Ig class switch recombination by NF-kappaB: retroviral expression of RelB in activated B cells inhibits switching to IgG1, but not to IgE. Int Immunol. 2002; 14(9):983-991. (Clone-specific: Flow cytometry). View Reference
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Honjo T, Obata M, Yamawaki-Katoaka Y, et al. Cloning and complete nucleotide sequence of mouse immunoglobulin gamma 1 chain gene. Cell. 1979; 18(2):559-568. (Biology). View Reference
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Ozaki K, Spolski R, Ettinger R, et al. Regulation of B cell differentiation and plasma cell generation by IL-21, a novel inducer of Blimp-1 and Bcl-6. J Immunol. 2004; 173(9):5361-5371. (Clone-specific: Flow cytometry). View Reference
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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.