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Ultrapure formaldehyde

Manufactured by Tousimis
Sourced in United States

Ultrapure formaldehyde is a high-purity chemical compound used as a laboratory reagent. It is a colorless, flammable gas that is soluble in water and other polar solvents. Ultrapure formaldehyde is typically used in various scientific and research applications that require a high degree of purity and consistency.

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2 protocols using ultrapure formaldehyde

1

Rhesus Macaque Whole Blood Phenotyping

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A 22-color panel was designed to examine the changes in cell phenotypes in whole blood collected from animals during the course of the study (surface staining for CD3, CD4, CD8 (two clones), CD11b, CD11c, CD14, CD16, CD20, CD25, CD28, CD45, CD49d, CD56, CD66abce, CD95, CD123, CD127, CD159 (NKG2a), CD194 (CCR4), and HLA-DR). All antibodies were selected based on cross-reactivity with rhesus macaques and fluorochrome availability. Antibody information, including clones and fluorochromes, is listed in Table S2.
Briefly, 100 μL of fresh EDTA whole blood was stained with Live/Dead Fixable Blue dye (ThermoFisher Scientific) and surface-stained at room temperature for 30 min. BD FACS Lysing solution (BD Biosciences, San Jose, CA, USA) was then used to lyse red blood cells according to the manufacturer’s instructions. Samples were washed twice with D-PBS and resuspended in 1% ultrapure formaldehyde (Tousimis, Rockville, MD, USA). Samples were acquired on a BD FACSymphony A5 analyzer using FACSDiva 8 software. Data were analyzed by using the FlowJo (version 10.6.) gating strategy as outlined in Figure S1A,B, established by using a combination of isotype and fluorescence-minus-one (FMO) controls.
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2

Multicolor Flow Cytometry Analysis of Macaque PBMCs

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Two staining panels were developed for analysis of macaque PBMC from these experiments. A 22-color panel was designed to examine T-cell and NK cell responses to treatment (CD2, CD3, CD4, CD7, CD8 [2 clones], CD14, CD16, CD20, CD56, NKp44, NKG2A, CD45, CD25, CD62L, CD28, CD95, CCR4, CD107a, CD127, and HLA-DR). A 16-color panel was designed to measure monocyte responses to treatment (CD3, CD4, CD11b, CD14, CD15, CD16, CD20, CD33, CD45, CD68, CD86, CCR2, CXCR4, PDL1, and HLA-DR). All antibodies were selected based on cross-reactivity with rhesus macaques and fluorochrome availability. Antibody information for both panels, including clones and fluorochromes, is listed in S3 Table. Frozen PBMCs were thawed, counted, and stained for flow cytometry analysis. Live/Dead Fixable aqua dye (Thermo Fisher Scientific) was included in each panel to eliminate dead cells.
Briefly, frozen PBMCs were thawed, counted, and resuspended at 1x107 cells/ml in D-PBS without Ca2+ or Mg2+ (Thermo Fisher Scientific). Cells were aliquoted into three 12 x 75 mm polystyrene tubes, 100 μl per tube, (Unstained, 22-color panel, 16-color panel) and stained for 30 min on ice. Samples were washed with D-PBS and resuspended in 1% ultrapure formaldehyde (Tousimis, Rockville, MD). Samples were acquired immediately on a BD FACSymphony A5 analyzer using FACSDiva 8 software. Data were analyzed using FlowJo Version 10.6.
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