PE Mouse Anti-Human IgG2

BD Pharmingen™ PE Mouse Anti-Human IgG2

Name: PE Mouse Anti-Human IgG2
Brand: BD Pharmingen™
SKU: 568960

Clone HP6002 (also known as AB4-DH4)

(RUO)

Multicolor flow cytometric analysis of IgG2 expression on Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were washed and cultured in complete tissue culture medium overnight to minimize subsequent 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 IgG2 antibody (Cat. No. 568959/568960; Right Plot). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of cell surface IgG2 (or Ig isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

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Product Details

Brand: BD Pharmingen™

Alternative Name: IGHG2; Immunoglobulin heavy constant gamma 2

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Purified Human IgG2 Myeloma Protein

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl/test

Entrez Gene ID: 3501

Storage Buffer: Aqueous buffered solution containing BSA, protein stabilizer, 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.
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.
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).
Source of all serum proteins is from USDA inspected abattoirs located in the United States.

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

DAPI Solution RUO

Size 1 mg Cat No. 564907

568960 Rev. 3

Antibody Details

HP6002

The HP6002 monoclonal antibody specifically recognizes the subclass of human Immunoglobulin G (IgG) known as IgG2. After human IgG1, human IgG2 is the second most abundant of the four subclasses of IgG found in human serum followed by IgG3 and IgG4. IgG2 is comprised of two identical heavy chains encoded by IGHG2, and two light chains, either Igκ or Igλ, linked by disulfide bonds. The HP6002 antibody binds to the CH2 domain of the IgG2 heavy chain and does not crossreact with other immunoglobulin heavy chain (IgH) subclasses. IgG2 is normally expressed by plasmablasts, plasma cells, and memory B cells as well as by some myeloma or plasmacytoma cells. HP6002 binds to soluble human IgG2, cytophilic IgG2 attached to cells through its Fc region, or human IgG2 antibodies specifically bound to antigens. IgG2 can cross the placenta and disseminate throughout extravascular fluids throughout the body. Human IgG2 serves multiple functions with the transmembrane form serving as an antigen receptor for B lymphocytes and secreted soluble forms participating in various effector functions including antibody-dependent neutralization of toxins or infection by microbes and complement fixation.

568960 Rev. 3

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

568960 Rev.3

Citations & References

View product citations for antibody "568960" on CiteAb

Development References (5)

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: Immunofluorescence, Immunoprecipitation). 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. (Clone-specific: Flow cytometry). View Reference

View All (5)

568960 Rev. 3

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.