PE Mouse Anti-Human IgG3

BD Pharmingen™ PE Mouse Anti-Human IgG3

Name: PE Mouse Anti-Human IgG3
Brand: BD Pharmingen™
SKU: 568297

Clone HP6050 (also known as CC4-DC10)

(RUO)

Two-color flow cytometric analysis of IgG3 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 IgG1, κ Isotype Control (Cat. No. 349043; Left Plot) or PE Mouse Anti-Human IgG3 antibody (Cat. No. 568296; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of cell surface IgG3 (or Ig isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

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Product Details

Brand: BD Pharmingen™

Alternative Name: IGHG3; Ig gamma-3 chain C region; Immunoglobulin heavy constant gamma 3

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Human IgG3 myeloma protein

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: 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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.

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).
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.
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).

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

APC Mouse Anti-Human CD19 RUO

Size 100 Tests Cat No. 555415

PE Mouse IgG1, κ Isotype Control RUO (GMP)

Size 100 Tests Cat No. 349043

568297 Rev. 2

Antibody Details

HP6050

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.

568297 Rev. 2

Format Details

R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PE

Excitation Source: Yellow-Green 488 nm, 532 nm, 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

568297 Rev.2

Citations & References

View product citations for antibody "568297" on CiteAb

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):1767-1779. (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. 2017; In Press. (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

View All (6)

568297 Rev. 2

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.