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PerCP-Cy™5.5 Rat Anti-Mouse IgD
PerCP-Cy™5.5 Rat Anti-Mouse IgD
Two-color flow cytometric analysis of IgD expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Hamster Anti-Mouse CD3e antibody (Cat. No. 553066/561826) and either PerCp-Cy™5.5 Rat IgG2a, κ Isotype Control (Cat. No. 550765; Left Panel) or PerCP-Cy™5.5 Rat Anti-Mouse IgD antibody (Cat. No. 564273, Right Panel). Two-color contour plots showing the correlated expression of IgD (or Ig isotype control staining) and CD3e were derived from gated events with the forward and side light scattering characteristics of viable splenocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Two-color flow cytometric analysis of IgD expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Hamster Anti-Mouse CD3e antibody (Cat. No. 553066/561826) and either PerCp-Cy™5.5 Rat IgG2a, κ Isotype Control (Cat. No. 550765; Left Panel) or PerCP-Cy™5.5 Rat Anti-Mouse IgD antibody (Cat. No. 564273, Right Panel). Two-color contour plots showing the correlated expression of IgD (or Ig isotype control staining) and CD3e were derived from gated events with the forward and side light scattering characteristics of viable splenocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
IGHD; Igh-5; Immunoglobulin heavy chain 5; Ig delta chain C region
Mouse (QC Testing)
Rat IgG2a, κ
Not reported
Flow cytometry (Routinely Tested)
0.2 mg/ml
380797
AB_2738722
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

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. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  5. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  6. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Cy is a trademark of GE Healthcare.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564273 Rev. 2
Antibody Details
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11-26c.2a

The 11-26c.2a monoclonal antibody specifically binds to mouse immunoglobulin D of all Igh-C haplotypes (e.g., IgDa, IgDb, IgDe), and it does not react with other immunoglobulin isotypes.  Although 11-26c.2a mAb binds membrane IgD expressed on the splenic B-cell surface with high affinity, it does not induce proliferation of splenic B cells in vitro.  In vivo injection of 11-26c.2a antibody does not have any effect on activation of mature B cells, as determined by MHC class II antigen expression.

564273 Rev. 2
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
564273 Rev.2
Citations & References
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Development References (4)

  1. Campbell KS, Cambier JC. B lymphocyte antigen receptors (mIg) are non-covalently associated with a disulfide linked, inducibly phosphorylated glycoprotein complex. EMBO J. 1990; 9(2):441-448. (Clone-specific: Immunoprecipitation). View Reference
  2. Hamilton AM, Lehuen A, Kearney JF. Immunofluorescence analysis of B-1 cell ontogeny in the mouse. Int Immunol. 1994; 6(3):355-361. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
  3. Ishihara K, Wood WJ Jr, Wall R, et al. Multiple B29 containing complexes on murine B lymphocytes. Common and stage-restricted Ig-associated polypeptide chains. J Immunol. 1993; 150(6):2253-2262. (Clone-specific: Flow cytometry). View Reference
  4. Nitschke L, Kosco MH, Kohler G, Lamers MC. Immunoglobulin D-deficient mice can mount normal immune responses to thymus-independent and -dependent antigens. Proc Natl Acad Sci U S A. 1993; 90(5):1887-1891. (Clone-specific: Flow cytometry). View Reference
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564273 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.