BD Horizon™ RY775 Mouse Anti-Human IgG
Clone G18-145 (RUO)
Multicolor flow cytometric analysis of IgG expression on viable Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were cultured in complete tissue culture medium overnight, washed, and then preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) to minimize subsequent nonspecific immunofluorescent staining. The cells were stained with BD Horizon™ BV421 Mouse Anti-Human CD19 antibody (Cat. No. 562440) and with either BD Horizon™ RY775 Mouse IgG1, κ Isotype Control (Cat. No. 571408; Left Plot) or BD Horizon™ RY775 Mouse Anti-Human IgG antibody (Cat. No. 571641/571720; Right Plot). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of IgG1 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
Multicolor flow cytometric analysis of IgG expression on viable Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were cultured in complete tissue culture medium overnight, washed, and then preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) to minimize subsequent nonspecific immunofluorescent staining. The cells were stained with BD Horizon™ BV421 Mouse Anti-Human CD19 antibody (Cat. No. 562440) and with either BD Horizon™ RY775 Mouse IgG1, κ Isotype Control (Cat. No. 571408; Left Plot) or BD Horizon™ RY775 Mouse Anti-Human IgG antibody (Cat. No. 571641/571720; Right Plot). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of IgG1 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
Product Details
BD Horizon™
IGHG1, IGHG2, IGHG3 and IGHG4
Human (QC Testing)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl/test
3500, 3501, 3502, 3503
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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.
- 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.
- 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.
- 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).
- 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 refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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.
- 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.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. 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 the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RY775 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
571408
BV421 Mouse Anti-Human CD19 RUO
Size
100 Tests
Cat No.
562440
Cell Viability Solution RUO
Size
500 Tests
Cat No.
555815
Human BD Fc Block™ RUO
Size
50 µg
Cat No.
564219
571641 Rev. 1
Antibody Details
G18-145
IgG is an important component of the humoral immune response, helping to control infection. IgG is produced by plasma B-cells and may be found in extracellular fluids, such as blood, lymph, peritoneal, and cerebrospinal fluids. IgG monomers consist of two light and two heavy chains containing two antigen binding sites. There are four IgG subclasses found in human, mouse and rat species, which include IgG1, IgG2, IgG3 and IgG4. The G18-145 monoclonal antibody specifically binds to the heavy chain of human immunoglobulin G subclasses: IgG1, IgG2, IgG3 and IgG4. The G18-145 antibody has been reported not to react with the heavy chains of other human immunoglobulin isotypes.
571641 Rev. 1
Format Details
RY775
The BD Horizon RealYellow™ 775 (RY775) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 775-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY775 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY775 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter).
RY775
Yellow-Green 561 nm
557 nm
775 nm
571641 Rev.1
Citations & References
View product citations for antibody "571641" on CiteAb
Development References (4)
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Jourdan M, Caraux A, Caron G, et al. Characterization of a transitional preplasmablast population in the process of human B cell to plasma cell differentiation. J Immunol. 2011; 187(8):3931-3941. (Clone-specific: Flow cytometry). View Reference
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Odendahl M, Mei H, Hoyer BF, et al. Generation of migratory antigen-specific plasma blasts and mobilization of resident plasma cells in a secondary immune response. Blood. 2005; 105(4):1614-1621. (Clone-specific: Flow cytometry). View Reference
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Roll P, Palanichamy A, Kneitz C, Dorner T, Tony HP. Regeneration of B cell subsets after transient B cell depletion using anti-CD20 antibodies in rheumatoid arthritis. Arthritis Rheum. 2006; 54(8):2377-2386. (Clone-specific: Flow cytometry). View Reference
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Scheeren FA, Naspetti M, Diehl S, et al. STAT5 regulates the self-renewal capacity and differentiation of human memory B cells and controls Bcl-6 expression. Nat Immunol. 2005; 6(3):303-313. (Clone-specific: Flow cytometry). View Reference
571641 Rev. 1
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.