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APC Rat Anti-Mouse IgM
APC Rat Anti-Mouse IgM

Multicolor flow cytometric analysis of mouse IgM expression on mouse splenocytes. C57BL/6 mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with PE Hamster Anti-Mouse CD3e (Cat. No. 561824/553063/553064) and either APC Rat IgG2a κ Isotype Control (Cat. No. 553932; Left Plot) or APC Rat Anti-Mouse IgM antibody (Cat. No. 550676/560232; Right Plot) at 0.25 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of IgM (or Ig Isotype control) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer Sy553stem and FlowJo™ software.

Multicolor flow cytometric analysis of mouse IgM expression on mouse splenocytes. C57BL/6 mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with PE Hamster Anti-Mouse CD3e (Cat. No. 561824/553063/553064) and either APC Rat IgG2a κ Isotype Control (Cat. No. 553932; Left Plot) or APC Rat Anti-Mouse IgM antibody (Cat. No. 550676/560232; Right Plot) at 0.25 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of IgM (or Ig Isotype control) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer Sy553stem and FlowJo™ software.

Product Details
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BD Pharmingen™
Mouse (QC Testing)
Rat IgG2a, κ
Not reported
Flow cytometry (Routinely Tested)
0.2 mg/ml
16019
AB_398464
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 and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

For flow cytometric detection of intracytoplasmic IgM, the use of FITC-conjugated rat anti-mouse IgM (clone II/41) (Cat. No. 553437) is recommended.

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

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
562032 Rev. 7
Antibody Details
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II/41

The II/41 clone has been reported to react specifically with mouse IgM of Igh-C[a] and Igh-C[b] haplotypes. It has been reported not to react with other Ig isotypes. In addition, the II/41 clone has been reported not to stimulate B-cell proliferation.

562032 Rev. 7
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC
Red 627-640 nm
651 nm
660 nm
562032 Rev.7
Citations & References
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Development References (2)

  1. Hennet T, Chui D, Paulson JC, Marth JD. Immune regulation by the ST6Gal sialyltransferase.. Proc Natl Acad Sci USA. 1998; 95(8):4504-9. (Clone-specific: Flow cytometry). View Reference
  2. Laszlo G, Hathcock KS, Dickler HB, Hodes RJ. Characterization of a novel cell-surface molecule expressed on subpopulations of activated T and B cells. J Immunol. 1993; 150(12):5252-5262. (Clone-specific). View Reference
562032 Rev. 7

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.