Tetraethylorthosilicate (TEOS), and polyethyleneimine (PEI) were obtained from Macklin Biochemical Technology Co., Ltd. (Shanghai, China). Ammonia water (NH3·H2O, 14.84 M), triethanolamine, 3-triethoxysilylpropylamine (APTE), anhydrous sodium carbonate (Na2CO3), curcumin (Cur), glutaraldehyde (GA), and phosphate buffered saline (PBS), were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Dialysis membrane and cetyltrimethylammonium bromide (CTAB) were obtained from Sangon Biotech Co., Ltd. (Shanghai, China). Tryptone, yeast extract was obtained from Oxoid Ltd. (Basingstoke, New Hampshire, UK). Agar was obtained from Beijing Jin Ming Biotechnology Co., Ltd. (Beijing, China) Trypticase Soy Broth was obtained from Qingdao Hope Bio-Technology Co., Ltd. (Qindao, China) S. aureus (ATCC29213) is a laboratory collection standard strain. All other chemicals in the experiment are analytical reagents and can be used without further purification.
Dialysis membrane
Manufactured by Sangon
Sourced in China
Dialysis membrane is a semi-permeable membrane used in various laboratory applications. Its core function is to allow the selective passage of small molecules while retaining larger molecules or particles. This membrane is commonly used in dialysis procedures to separate and purify substances based on their molecular size.
Automatically generated - may contain errors
Lab products found in correlation
Phosphate buffered saline (pbs), by Sinopharm (1 mentions)
Trypticase soy broth (tsb), by Hopebio (1 mentions)
Tryptone yeast extract, by Thermo Fisher Scientific (1 mentions)
Tetraethylorthosilicate teos, by Maclin Biochemical Technology (1 mentions)
Polyethyleneimine pei, by Maclin Biochemical Technology (1 mentions)
Ni nta 6ff, by Sangon (1 mentions)
Axyprep plasmid miniprep kit, by Corning (1 mentions)
Bca protein quantification kit, by Vazyme (1 mentions)
Hrp conjugated goat anti mouse igg, by Huabio (1 mentions)
Ni nta sefinose resin, by Sangon (1 mentions)
Imidazole, by Sangon (1 mentions)
Hrp conjugated goat anti rabbit igg, by Huabio (1 mentions)
T4 dna ligase, by Vazyme (1 mentions)
Urea, by Sangon (1 mentions)
4 protocols using dialysis membrane
1
TEOS-PEI Nanoparticles for Curcumin Delivery
2
Cloning and Purification of XaffOBP9
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Specific primers were designed to clone the cDNA that encodes XaffOBP9 (Supplementary Table S1 ). The PCR products were inserted into the pET-28a (+) vector using NotI and NcoI restriction endonucleases. The plasmid containing the correct insert fragment was subsequently transformed into Escherichia coli BL21 (DE3) cells. The recombinant protein was induced at 28°C for 6 h by 1 mM isopropyl β-d-l-thiogalactopyranoside (IPTG) when the OD600 value reached 0.6. The suspension was sonicated and then separated into supernatant and sediment by centrifugation (11,000 rpm, 20 min, 4 C). The protein was then purified using Ni-NTA 6FF (Sangon Biotech, Shanghai, China) in a graded imidazole series of 0 mM, 20 mM, 40 mM, 60 mM, 80 mM, 100 mM, 200 mM, 400 mM, 600 mM for washing and desalted using Dialysis Membrane (Sangon Biotech, Shanghai, China). The molecular weight and purity of the XaffOBP9 proteins were checked using 15% SDS-PAGE.
3
DNA Extraction and Protein Purification Protocols
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DNA gel extraction kits (Omega Bio-tek, USA), AxyPrep plasmid miniprep kit (Axygen Scientific Inc, USA) were used for DNA fragments purification and plasmid extraction according to the manufacturer's instructions. The BCA protein quantification kit and T4 DNA ligase were from Vazyme Biotech (Nanjing, China). The following reagents were purchased from Sangon Biotech (Shanghai, China): urea, imidazole, Ni-NTA sefinose resin, dialysis membrane (cutoff MW, 14 KD), polyethylene glycol-20000 (PEG20000), 50 × TAE buffer, two-color pre-stained protein marker. The 2 × Hieff PCR Master mixture, nucleic acid gel stain (10000 × in water), and DNA marker were obtained from Yeasen Biotech (Shanghai, China). NcmECL Ultra Kit was obtained from NCM Biotech (Suzhou, China) and used for detection of Western blot.
The mouse anti-6 × His tag monoclonal antibodies was from Proteintech Group (USA). The rabbit polyclonal antibody against SARS-CoV2 Spike RBD was from ABclonal Technology (Wuhan, China). Antibodies against the following agents were obtained from BBI Life Sciences (Shanghai, China): rabbit anti-ACE2 polyclonal antibody, rabbit anti-Myc tag polyclonal antibody, HRP-conjugated goat anti-rabbit IgG, HRP-conjugated goat anti-mouse IgG. Goat anti-mouse IgG-Alexa Fluor® 647 antibody was purchased from HuaBio (Hangzhou, China).
The mouse anti-6 × His tag monoclonal antibodies was from Proteintech Group (USA). The rabbit polyclonal antibody against SARS-CoV2 Spike RBD was from ABclonal Technology (Wuhan, China). Antibodies against the following agents were obtained from BBI Life Sciences (Shanghai, China): rabbit anti-ACE2 polyclonal antibody, rabbit anti-Myc tag polyclonal antibody, HRP-conjugated goat anti-rabbit IgG, HRP-conjugated goat anti-mouse IgG. Goat anti-mouse IgG-Alexa Fluor® 647 antibody was purchased from HuaBio (Hangzhou, China).
4
Quantifying Microcystis Exopolysaccharide Production
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The growth of M. aeruginosa strains was determined every two days by measuring the dry cell weight. The M. aeruginosa liquid culture samples (100 ml) were centrifuged at 7,000 × g for 10 min. Then cells were harvested, washed three times with distilled water, and finally dried at 105 °C to a constant weight for measuring the dry weight. At the same time, the supernatants were collected and concentrated to about 40 ml by a rotary evaporator. Then powder active carbon was added for depigmentation and was removed thereafter by passing through a 0.45-µm filtration membrane. The clarified liquor was mixed with 4 times the volume of 95% ethanol and incubated overnight at 4 °C for EPS precipitation. The precipitate was concentrated by evaporation and resuspended with distilled water, and this was repeated twice to completely remove the ethanol. Then the EPS collection was dried at 60 °C for dry weight measurement. The prepared EPS was dissolved in distilled water (0.01%, w/v) as a stock solution for further tests. The polysaccharide solution was then applied to a dialysis membrane for the separation of 1, 3.5, and 5 kDa molecule (Sangon Biotech Co., Ltd., Shanghai). The concentration of each of the dialyzed fractions was determined by the phenol–sulfuric acid method43 (link), using glucose as the standard.
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