Ultrapure sds
Manufactured by Thermo Fisher Scientific
UltraPure SDS is a laboratory reagent used for the preparation of sodium dodecyl sulfate (SDS) solutions. SDS is a commonly used anionic detergent in various applications, such as protein electrophoresis and Western blotting. UltraPure SDS is designed to provide a high-purity, low-residue solution for consistent and reliable results in these applications.
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Lab products found in correlation
2 protocols using ultrapure sds
1
Endotoxin-free Bacillus anthracis PGN Purification
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Bacillus anthracis Delta Sterne PGN was prepared using nonpyrogenic labware and endotoxin-free water for all solutions as previously described (8 (link), 9 (link)). Briefly, four 500-ml cultures of vegetative bacteria were collected after overnight growth in Trypticase soy broth and subsequently washed once with endotoxin-free water. The cultures were collected by centrifugation at 15,000 × g for 10 min at 4°C and then resuspended and boiled three times in 8% UltraPure SDS (Invitrogen). After the final boil, the material was treated twice with DNase/RNase for 30 min at room temperature, followed by washes with endotoxin-free water and centrifugation as described above. Hydrofluoric acid was added to the material overnight at 4°C in order to remove any teichoic acid residues. Following hydrofluoric acid treatment, the material was washed and resuspended in proteinase K solution overnight at 50°C. Proteinase K digestion was conducted twice, followed by washing and subsequent sonication. The now-purified PGN was then boiled in 8% SDS, washed, and dried. PGN was weighed and resuspended in endotoxin-free water at 20 mg/ml.
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2
MTT Assay for Cell Proliferation
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The tetrazolium bromide (MTT) assay was used to determine cell proliferation for 4–7 days. Cells were plated after 5 days of LV transduction and 3 days of puromycin selection, except for SUDHL4 cells, which were plated 3 days after LV transduction. Briefly, 100 μL of cells were seeded in 96-well plates at a concentration of 5k/well for primary dermal fibroblasts, 20k/well for HBL1, HL60, K562; 40k/well for U2932, OCI-Ly7, and SUDHL4; 3k/well for SNU16, SW480, and LS1034. On each consecutive day, cells were incubated with 5 mg/mL MTT for 3 hours at 37°C. The reaction was stopped by the addition of 100 μL of 10% Ultrapure SDS (Invitrogen) to dissolve precipitated crystals. After overnight incubation at 37°C, the absorbance was measured on a microplate spectrophotometer at 570 nm while subtracting the background absorbance measured at 690 nm.
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