Whatman filter paper #1 was purchased from GE Healthcare Life Sciences. Wax paper squares (6″) were purchased from NORPRO. Gold (III) chloride hydrate, sodium citrate tribasic, poly (ethylene glycol) 2-mercaptoethyl ether acetic acid (thiol-PEG-acid) 2100, N-hydroxysulfosuccinimide sodium salt (sulfo-NHS), N-(3-Dimethylaminopropyl)-N′- ethylcarbodiimide hydrochloride (EDAC; EDC), poly (sodium 4-styrenesulfonate, average Mw ∼70.000) 30 % solution, polyethylene glycol 3000 (biotin-PEG), Tween-20, 2-ethanesulfonic acid (MES), and anti-mouse IgG (Fc Specific) (SAB3701021) were obtained from Sigma Aldrich. Mouse monoclonal antibody Anti-SARS-CoV-2 N-protein (35–579) was purchased from ProScience. Recombinant Nucleocapsid protein from SARS-CoV-2 (230−01104) was purchased from Raybiotech. Bovine serum albumin (BSA, protease free) was purchased from VWR chemicals. Aerosol bottles were purchased from Lurrose. Surgical facemasks for experiments in Fig. 3 were purchased from Medline (NONE27377). PBS refers to phosphate buffered saline pH 7.4. PBST refers to PBS supplemented with 0.1 % Tween-20. RT refers to room temperature. PBS-BSA refers to PBS supplemented with 0.5 % BSA.
2 ethanesulfonic acid mes
Manufactured by Merck Group
2-ethanesulfonic acid (MES) is a chemical compound used as a buffer in biological applications. It is a zwitterionic, Good's buffer that maintains a stable pH in the range of 5.5 to 6.7. MES is commonly used to maintain pH in cell culture media, biochemical assays, and other laboratory procedures where pH control is critical.
Automatically generated - may contain errors
Lab products found in correlation
Gold 3 chloride hydrate, by Merck Group (2 mentions)
Sodium citrate tribasic, by Merck Group (1 mentions)
Whatman 1 filter paper, by GE Healthcare (1 mentions)
Ab6672, by Abcam (1 mentions)
Sodium citrate tribasic dihydrate, by Merck Group (1 mentions)
N hydroxysulfosuccinimide sodium salt sulfo nhs, by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
Peroxidase from horseradish, by Merck Group (1 mentions)
Observer z1 lsm700 confocal microscope, by Zeiss (1 mentions)
Bx50wi, by Olympus (1 mentions)
3 3 diaminobenzidine (dab), by Merck Group (1 mentions)
Alexafluor 647 phalloidin, by Merck Group (1 mentions)
Fluoromount g, by Electron Microscopy Sciences (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Glycine, by Merck Group (1 mentions)
4 protocols using 2 ethanesulfonic acid mes
1
Development of SARS-CoV-2 Antigen Assay
2
Fabrication of IL-6 Immunosensor
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whatman filter paper #41 and #1 was obtained from GE Healthcare Life Sciences. Wax Paper Squares (6″) was purchased from NORPRO. Gold (III) chloride hydrate, sodium citrate tribasic dihydrate, poly (ethylene glycol) 2-mercaptoethyl ether acetic acid (thiol-PEG-acid) 2100, N-Hydroxysulfosuccinimide sodium salt (sulfo-NHS), N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDAC; EDC), O-(2-biotinyl-aminoethyl) polyethylene glycol 3000 (biotin-PEG), poly (sodium 4-styrenesulfonate, average Mw ≈ 70.000) 30 % solution, Tween-20, 2-ethanesulfonic acid (MES) and recombinant human IL-6 (I2786) were obtained from Sigma Aldrich. Rabbit anti-IL-6 polyclonal antibody (ab6672) was purchased from Abcam. Albumin from bovine serum (BSA, protease free) was obtained from VWR Chemicals. PBS refers to phosphate buffered saline pH 7.4. PBST refers to PBS supplemented with 0.1 % Tween-20. RT refers to room temperature.
3
Visualizing Subcellular Hydrogen Peroxide
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DAB staining: DAB is oxidized by H2O2 in the presence of peroxidases to generate a dark brown precipitate (Thordal-Christensen et al., 1997 ). Seedlings were incubated with 1 mg·mL−1 DAB (Sigma-Aldrich) in 20-mM 2-ethanesulfonic acid (MES; Sigma-Aldrich) buffer (pH 6.2) supplemented by 10 units·mL−1 peroxidase from horseradish (Sigma-Aldrich) for 1 h. Seedlings were then rinsed with MES buffer for 1 min twice before imaging (Dubreuil-Maurizi et al., 2011 (link)). Olympus BX50WI was used to visualize DAB staining and images were taken under 10× objective lens.
Confocal imaging of mt-roGFP2-Orp1: to visualize subcellular levels of H2O2, confocal imaging of 5-day-old mt-roGFP2-Orp1 Arabidopsis seedlings was performed right after experimental time-points with a Zeiss Observer Z1 LSM700 confocal microscope (oil immersion, 40× objective Plan-Neofluar N.A. 1.30). RoGFP2-Orp1 was excited sequentially at 405 and 488 nm and emission was recorded at 505–535 nm. The 405/488 ratio within root tips was determined in similar areas of ∼5,000 μm2 using ICY software (http://icy.bioimageanalysis.org ).
Confocal imaging of mt-roGFP2-Orp1: to visualize subcellular levels of H2O2, confocal imaging of 5-day-old mt-roGFP2-Orp1 Arabidopsis seedlings was performed right after experimental time-points with a Zeiss Observer Z1 LSM700 confocal microscope (oil immersion, 40× objective Plan-Neofluar N.A. 1.30). RoGFP2-Orp1 was excited sequentially at 405 and 488 nm and emission was recorded at 505–535 nm. The 405/488 ratio within root tips was determined in similar areas of ∼5,000 μm2 using ICY software (
4
Fixation and Fluorescence Staining of Adherent Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RPE1 vin-eGFP and NIH-3T3 vin-eGFP cells were fixed after 5 hours of spreading. First, cells were prepermeabilized using 0.25% Triton X-100 (Sigma-Aldrich) diluted in cytoskeleton buffer [10 mM 2-ethanesulfonic acid (MES, Sigma-Aldrich), 100 mM potassium chloride (KCl, Sigma-Aldrich), and 3.6 mM magnesium chloride hexahydrate (MgCl 6H2O, Sigma-Aldrich), and 1.9 mM aminopolycarboxylic acid (EGTA, Sigma-Aldrich) in ddH2O]. The sample was quickly rinsed with the prepolymerization solution and immediately placed into the fixation buffer [4% paraformaldehyde and 10% (w/v) sucrose (Sigma-Aldrich) in cytoskeleton buffer]. After 15-min fixation at RT, samples were washed once with cytoskeleton buffer and blocked for 30 min at RT with blocking buffer [0.5% bovine serum albumin (Sigma-Aldrich), 0.1% sodium azide (NaN3, Sigma-Aldrich), and 20 mM glycine (Sigma-Aldrich) in dPBS]. For additional staining of filamentous actin (F-actin), samples could be incubated for 30 min at RT with Alexa Fluor 647 phalloidin (Sigma-Aldrich) diluted in blocking buffer (1:1000). The fixed sample was mounted onto a glass slide using Fluoromount-G (Electron Microscopy Sciences) and stored at 4°C.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!