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.
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. For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
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 to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. 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.
2. Researchers should determine the optimal concentration of this reagent for their individual applications.
3. An isotype control should be used at the same concentration as the antibody of interest.
4. 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.
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. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
9. 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.
10. CF™ is a trademark of Biotium, Inc.
11. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
12. BD Horizon Brilliant Violet 605 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
13. 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.
