Silicon nitride AFM cantilever, PNP-TR (NanoWorld, Neuchâtel, Switzerland), was cleaned by sonication in a series of solvents and functionalization in 3-aminopropyl-trimethoxysilane and glutaraldehyde (Grade II, Sigma) according to the method reported previously with minor modifications25 (link), 26 (link), 55 (link). For anchoring proteins, the tips were rinsed with PBS to remove the glutaraldehyde remainders and subsequently incubated with proteins, including BSA, Pam3CSK4, rLipL32 and its variants for 1 h at room temperature. The functionalized AFM tips were then rinsed three times with PBS to remove the unbound proteins56 (link). The mica surface was modified for better deposition of proteins according to previous reports57 (link), 58 (link). The unbound proteins were removed and the proteins were fixed on the mica for later AFM measurements.
Grade 2
Manufactured by Merck Group
Sourced in Switzerland, Italy
Grade II is a line of specialized laboratory equipment produced by Merck Group. It is designed for precise and efficient performance of various scientific tasks in research and testing environments. The core function of this product line is to provide reliable and high-quality tools for laboratory operations.
Automatically generated - may contain errors
Lab products found in correlation
Grade 5, by Merck Group (2 mentions)
Pnp tr, by NanoWorld (1 mentions)
Accuri c6, by BD (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
Dulbecco s phosphate buffered saline, by Merck Group (1 mentions)
Aluminum hydroxide, by Merck Group (1 mentions)
Ovalbumin (ova), by Merck Group (1 mentions)
4 protocols using grade 2
1
Functionalization of AFM Cantilevers for Protein Attachment
2
Murine Allergic Asthma Model with Dexamethasone
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The protocol of experimental model establishing was adopted as previously described [12 (link)]. Mice were sensitized IP with 20 μg chicken OVA (Grade V, Sigma Aldrich Co., St. Louis, MO) emulsified in 2.0 mg of alum (Shanghai No.4 Reagent & H.V. Chemical Industries Ltd., Shanghai, China) in a total volume of 100 μl of 0.9% sterile saline on days 1 and 14. Non-sensitized mice just received 2.0 mg of alum in 0.9% saline. On day 24, 25, and 26, mice received three aerosol challenges with 5% OVA (wt/vol) (Grade II, Sigma Aldrich Co., St. Louis, MO) in 0.9% endotoxin-free saline (non-sensitized mice received saline only) for 30 min daily. Using a plastic box (50 × 30 × 40 cm3) as exposure chamber, where the air was kept at 20 – 22°C and relative humidity 50 – 60%, the aerosol was generated by a sidestream jet nebulizer (PARI BOY, Starnberg, Germany) into the chamber and mice were put into stainless cages inside the chamber with irradiated food and acidified water provided ad libitum. One hour before each challenge, mice received dexamethasone (2 mg/kg) IP in 0.1 ml endotoxin-free PBS or PBS alone. The dosage of dexamethasone has been adopted by multiple publications [16 (link)].
3
Peat Water Microbial Abundance Quantification
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For each layer of each sampling point, at least 20 mL of peat water was extracted, prefiltered at 125 μm, and fixed in Glutaraldehyde (2%, Grade II, Sigma-Aldrich). Prokaryote abundance (PA) and viral particle abundance (VPA) were obtained using flow cytometry (BD Accuri C6) (Sime-Ngando et al., 2008 (link)).
4
Ozone Exposure in Ovalbumin-Induced Asthma Model
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C57BL/6 mice were randomly divided into three groups with eight mice in each group (see schematic diagram of mouse experiment in Figure S2
Mice were exposed to ozone at a concentration of 3 ppm, or air in a Perspex container 30 min after each OVA/PBS challenge on days 14, 16, 18, 20, and 22. Ozone was produced by an ozone generator (model 300 Ozoniser, Aqua Medic, Germany). Every exposure lasted 2 h. Ozone concentration was continuously controlled and adjusted by an OS-4 ozone switch (Eco Sensors, KWJ Engineering Inc, California, USA). The control mice were exposed to air during this time.
Mice were exposed to ozone at a concentration of 3 ppm, or air in a Perspex container 30 min after each OVA/PBS challenge on days 14, 16, 18, 20, and 22. Ozone was produced by an ozone generator (model 300 Ozoniser, Aqua Medic, Germany). Every exposure lasted 2 h. Ozone concentration was continuously controlled and adjusted by an OS-4 ozone switch (Eco Sensors, KWJ Engineering Inc, California, USA). The control mice were exposed to air during this time.
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