Preparation And Storage
Recommended Assay Procedures
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
- 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).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
Companion Products
The HP6050 monoclonal antibody specifically recognizes the subclass of human Immunoglobulin G (IgG) known as IgG3. Human IgG3 is the third most abundant of the four subclasses of IgG (IgG1, IgG2, IgG3, and IgG4) named in order of their relative serum concentrations. IgG3 is comprised of two identical heavy chains encoded by IGHG3, and two light chains, either Igκ or Igλ, linked by disulfide bonds. The HP6050 antibody binds to the hinge region of the IgG3 heavy chain and does not crossreact with other immunoglobulin heavy chain (IgH) subclasses. IgG3 is normally expressed by plasmablasts, plasma cells, and memory (IgG3 class-switched) B cells as well as by some myeloma or plasmacytoma cells. HP6050 binds to soluble human IgG3, cytophilic IgG3 attached to Fc receptor-positive cells through its Fc region, or human IgG3 antibodies specifically bound to antigens. IgG3 can cross the placenta and diffuse throughout extravascular fluids throughout the body. Human IgG3 serves multiple functions with the transmembrane form serving as an antigen receptor for B lymphocytes and secreted soluble forms participating in various effector functions. The latter include antibody-dependent neutralization of toxins or infection by microbes, opsonization for phagocytosis, efficient complement fixation, and cell-mediated cytotoxicity.
Development References (6)
-
Avery DT, Bryant VL, Ma CS, de Waal Malefyt R, Tangye SG. IL-21-induced isotype switching to IgG and IgA by human naive B cells is differentially regulated by IL-4.. J Immunol. 2008; 181(3):1767-79. (Biology). View Reference
-
Blanco E, Perez-Andres M, Sanoja-Flores L, et al. Selection and validation of antibody clones against IgG and IgA subclasses in switched memory B-cells and plasma cells.. J Immunol Methods. 2019; 475:112372. (Clone-specific: Flow cytometry). View Reference
-
Gao B, Moore C, Porcheray F, et al. Pretransplant IgG reactivity to apoptotic cells correlates with late kidney allograft loss.. Am J Transplant. 2014; 14(7):1581-91. (Clone-specific: Flow cytometry). View Reference
-
Jefferis R, Reimer CB, Skvaril F, et al. Evaluation of monoclonal antibodies having specificity for human IgG sub-classes: results of an IUIS/WHO collaborative study.. Immunol Lett. 1985; 10(3-4):223-52. (Clone-specific: ELISA, Hemagglutination assay, Immunocytochemistry, Immunofluorescence, Radioimmunoassay). View Reference
-
Reimer CB, Phillips DJ, Aloisio CH, et al. Evaluation of thirty-one mouse monoclonal antibodies to human IgG epitopes.. Hybridoma. 1984; 3(3):263-75. (Immunogen: Array, Fluorescence quantitation). View Reference
-
Tangye SG, Ferguson A, Avery DT, Ma CS, Hodgkin PD. Isotype switching by human B cells is division-associated and regulated by cytokines.. J Immunol. 2002; 169(8):4298-306. (Biology). View Reference
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.