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APC-H7 Rat Anti-Mouse IgD
APC-H7 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 BD Horizon™ PE-CF594 Hamster Anti-Mouse CD3e antibody (Cat. No. 562286/562332) and either APC-H7 Rat IgG2a, κ Isotype Control (Cat. No. 560197; Left Panel) or APC-H7 Rat Anti-Mouse IgD antibody (Cat. No. 565348; Right Panel). Two-color contour plots showing the correlated expression of IgD (or Ig isotype control staining) versus 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 BD Horizon™ PE-CF594 Hamster Anti-Mouse CD3e antibody (Cat. No. 562286/562332) and either APC-H7 Rat IgG2a, κ Isotype Control (Cat. No. 560197; Left Panel) or APC-H7 Rat Anti-Mouse IgD antibody (Cat. No. 565348; Right Panel). Two-color contour plots showing the correlated expression of IgD (or Ig isotype control staining) versus 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_2739201
Aqueous buffered solution containing protein stabilizer and ≤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 APC-H7 under optimum conditions, and unconjugated antibody and APC-H7 were removed.

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. BD APC-H7 is a tandem conjugate and an analog of APC-Cy7 with the same spectral properties. It has decreased intensity but it is engineered for greater stability and less spillover in the APC channel and consequently offers better performance than APC-Cy7. It has an absorption maximum of approximately 650 nm. When excited by light from a red laser, the APC fluorochrome can transfer energy to the cyanine dye, which then emits at a longer wavelength. The resulting fluorescent emission maximum is approximately 767 nm. BD recommends that a 750-nm longpass filter be used along with a red-sensitive detector such as the Hamamatsu R3896 PMT. As with APC-Cy7 special filters are required when using APC-H7 in conjunction with APC. Note: Although our APC-H7 products demonstrate higher lot-to lot consistency than other APC tandem conjugate products, and every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-H7 conjugate.
  6. Although BD APC-H7 is engineered to minimize spillover to the APC channel and is more stable and less affected by light, temperature, and formaldehyde-based fixatives, compared to other APC-cyanine tandem dyes, it is still good practice to minimize as much as possible, any light, temperature and fixative exposure when working with all fluorescent conjugates.
  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.
565348 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.

        

565348 Rev. 2
Format Details
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APC-H7
The BD Horizon™ APC-H7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 659 nm and an acceptor dye, H7, with an emission maximum (Em Max) at 782 nm. APC-H7, driven by BD innovation, is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC-H7
Red 627-640 nm
659 nm
782 nm
565348 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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565348 Rev. 2

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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.