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Mouse igg2b isotype control

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The Mouse IgG2b Isotype Control is a laboratory reagent used as a negative control in flow cytometry experiments. It is an antibody of the IgG2b subclass derived from mouse, and it does not bind to any specific target antigen. This control is used to establish the baseline level of non-specific binding in a flow cytometry assay, which helps distinguish specific from non-specific staining of cells.

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Lab products found in correlation

Nunc maxisorp, by Thermo Fisher Scientific (1 mentions) Glomax microplate reader, by Promega (1 mentions) Dihydrorhodamine 123, by Merck Group (1 mentions) Trypan blue, by Merck Group (1 mentions) Cd71 fitc, by Thermo Fisher Scientific (1 mentions) Protease phosphatase inhibitor, by Merck Group (1 mentions) Human m csf, by BioLegend (1 mentions) Cd11b pe, by Thermo Fisher Scientific (1 mentions) Cd14 fitc, by Thermo Fisher Scientific (1 mentions) Cd68 fitc, by BioLegend (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Phorbol 12 myristate 13 acetate pma, by Abcam (1 mentions)

2 protocols using mouse igg2b isotype control

1

Quantification of Total DNA and MPO DNA

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For the total DNA assay, NET/DNA samples were stained with PicoGreen and fluorescence read on GloMax microplate reader (Promega, Madison, WI). The MPO/DNA ELISA was performed as previously described49 (link). Briefly, 96-well flat bottom Nunc MaxiSorp plates (Thermo Fisher Scientific) were coated with 5 μg/ml mouse IgG2b anti-human MPO (Clone: 4A4, Bio-Rad, Hercules, CA) or 5 μg/ml mouse IgG2b isotype control (Clone: 27–35, Biolegend) for 1 h at room temperature. Remaining steps were performed according to the Roche Cell Death Detection ELISA kit (Cat. No. 11544675001, Sigma-Aldrich).
Lelliott P.M., Hobro A.J., Pavillon N., Nishide M., Okita Y., Mizuno Y., Obata S., Nameki S., Yoshimura H., Kumanogoh A, & Smith N.I. (2023). Single-cell Raman microscopy with machine learning highlights distinct biochemical features of neutrophil extracellular traps and necrosis. Scientific Reports, 13, 10093.
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2

Characterization of Macrophage Phenotypes

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PE anti-human CD32 antibody, the mouse IgG2b isotype control, Human M-CSF, CD68-FITC, and the CD86-Alexa-Fluor antibodies were purchased from BioLegend (San Diego, CA). Benzo(a)pyrene [B(a)P] powder, Trypan blue, dimethyl sulfoxide (DMSO), dihydrorhodamine 123, and protease/phosphatase inhibitors were obtained from Sigma-Aldrich (St. Louis, MO). FITC-Dextran (Mr, 40,000 kDa) was purchased from Molecular Probes/Invitrogen (Carlsbad, CA). Phorbol 12-myristate 13-acetate (PMA) and Cholera Toxin Subunit B Hrp-conjugated ab (CTβ) were purchased from Abcam Biochemicals (Cambridge, MA). Specific antibodies to CD11b-PE, CD14-FITC, CD64, CD71-FITC, control IG1Κ and FITC were purchased from eBioscience (San Diego, CA). CD45 and CD59 were purchased from Santa Cruz Biotechnology (Dallas, TX). Amplex Red Cholesterol Assays and Anti-Human IgG(γ)-FITC conjugate ab were obtained from Invitrogen (Carlsbad, CA). Human IgG complexes (human heat-aggregated IgG; hHAIgG) were formed by incubating monomeric human IgG for 20 min at 63°C and centrifuged at 14,000 × g to removed precipitates.
Clark R.S., Pellom S.T., Booker B., Ramesh A., Zhang T., Shanker A., Maguire M., Juarez P.D., Matthews-Juarez P., Langston M.A., Lichtveld M.Y, & Hood D.B. (2016). Validation of Research Trajectory 1 of an Exposome Framework: Exposure to benzo(a)pyrene confers enhanced susceptibility to bacterial infection. Environmental research, 146, 173-184.
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