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Two-color flow cytometric analysis of IgG1 expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were washed and cultured in complete tissue culture medium overnight to minimize nonspecific immunofluorescent staining. The cells were harvested and stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415) and with either PE Mouse IgG2b, κ Isotype Control (Cat. No. 555058; Left Plot) or PE Mouse Anti-Human IgG1 antibody (Cat. No. 568275; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of cell surface IgG1 (or Ig isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ software.
BD Pharmingen™ PE Mouse Anti-Human IgG1
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
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 cell 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Companion Products
The HP6001 monoclonal antibody specifically recognizes the subclass of human Immunoglobulin G (IgG) known as IgG1. Human IgG1 is the most abundant of the four subclasses of IgG found in human serum followed by IgG2, IgG3 and IgG4. IgG1 is comprised of two identical heavy chains encoded by IGHG1 and two light chains, either Igκ or Igλ, linked by disulfide bonds. The HP6001 antibody binds to the CH2 domain of the IgG1 heavy chain and does not crossreact with other immunoglobulin heavy chain (IgH) subclasses. IgG1 is normally expressed by plasmablasts, plasma cells, and memory B cells as well as by some myeloma or plasmacytoma cells. HP6001 binds to soluble human IgG1, cytophilic IgG1 attached to cells through its Fc region, or human IgG1 antibodies specifically bound to antigens. IgG1 can cross the placenta and disseminate in extravascular fluids throughout the body. Human IgG1 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, complement fixation, and cell-mediated cytotoxicity.
Development References (7)
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Aucouturier P, Mounir S, Preud'homme JL. Distribution of IgG subclass levels in normal adult sera as determined by a competitive enzyme immunoassay using monoclonal antibodies. Diagn Immunol. 1985; 3(4):191-196. (Biology). View Reference
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Blanco E, Perez-Andres M, Arriba-Mendez S, et al. Age-associated distribution of normal B-cell and plasma cell subsets in peripheral blood. J Allergy Clin Immunol. 2018; 141(6):2208-2219. (Biology). View Reference
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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. 2017; In Press. (Clone-specific: Flow cytometry). View Reference
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Hamilton RG. Mohan C, ed. The Human IgG Subclasses. San Diego: Calbiochem-Novabiochem Corporation; 2001:1-64.
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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: Immunofluorescence). View Reference
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Liechti T, Günthard HF, Trkola A. OMIP-047: High-Dimensional phenotypic characterization of B cells.. Cytometry A. 2018; 93(6):592-596. (Clone-specific: Flow cytometry). View Reference
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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. (Clone-specific). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.