DNA sequences, hemin, and PDGF-BB (human) were from Sangon Biotechnology Inc. (Shanghai, People’s Republic of China). The Bst 2.0 DNA polymerase (large fragment), Nb.BbvCI nicking endonuclease, and deoxynucleotide (dNTP) solution mixture were obtained from New England Biolabs (Beijing, People’s Republic of China). Tris (2-carboxyethy) phosphine hydrochloride (TCEP) and dimethyl sulfoxide (DMSO) were purchased from Aladdin Reagent Corporation (Shanghai, People’s Republic of China). 6-Mercapto-1- hexanol (MCH) was obtained from J&K Scientific Ltd. (Guangzhou, People’s Republic of China). MCH solution was prepared by diluting the MCH with DNA hybridization buffer (20 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid [HEPES], 500 mM NaCl, pH 7.4) to 1 mM. The stock solution of hemin (10 mM) was prepared with DMSO and, then, stored at −20°C. DNA sequences were dissolved with 1× tris-EDTA (TE) buffer (pH 8.0) to 100 μM. Sequences of the DNA (Table 1 ) were designed with the help of the Integrated DNA Technologies (IDT; online software).
Hemin
Manufactured by Sangon
Sourced in China
Hemin is a chemical compound that serves as a cofactor for various enzymes involved in cellular processes. It is a key component of hemoglobin, the protein responsible for oxygen transport in red blood cells.
Automatically generated - may contain errors
Lab products found in correlation
Vitamin k1, by Sangon (3 mentions)
Peptone, by Merck Group (2 mentions)
Tryptone, by Merck Group (2 mentions)
Yeast extract, by Merck Group (2 mentions)
L cysteine hydrochloride, by Sangon (2 mentions)
Tween 80, by Merck Group (2 mentions)
Nb bbvci nicking endonuclease, by New England Biolabs (1 mentions)
Brain heart infusion broth, by Hopebio (1 mentions)
Phi29 dna polymerase, by Thermo Fisher Scientific (1 mentions)
Hydrogen peroxide h2o2, by Sangon (1 mentions)
Sulfuric acid h2so4, by Sangon (1 mentions)
Acetate, by Merck Group (1 mentions)
Propionate, by Merck Group (1 mentions)
Butyrate, by Merck Group (1 mentions)
Lactate, by Merck Group (1 mentions)
Succinate, by Merck Group (1 mentions)
Agar, by Sangon (1 mentions)
Dextran sulfate sodium (dss), by MP Biomedicals (1 mentions)
8 protocols using hemin
1
DNA-based Hemin-PDGF-BB Sensing Platform
2
Extraction and Characterization of Enteromorpha clathrata Polysaccharide
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Enteromorpha clathrata polysaccharide (ECP) was used and prepared as previously described [1 (link),5 (link)]. Tryptone, peptone, yeast extract and Tween 80 were obtained from Sigma (Shanghai, China). Hemin and L-cysteine hydrochloride were purchased from Sangon Biotech (Shanghai, China). All other chemicals of analytical grade were acquired from Sinopharm Chemical (Shanghai, China).
3
Isolation and Characterization of Bacteroides vulgatus Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We used B. vulgatus strains FTJS5K1 (5K1), FTJS7K1 (7K1), FSDTA11B14 (11B14), and FSDLZ51K1 (51K1) in this study, all of which had been isolated from the fecal samples of different volunteers. The 5K1 and 7K1 strains were deposited in the Culture Collection of Food Microorganisms (CCFM) of Jiangnan University (Wuxi, China). The four strains were grown anaerobically at 37°C for 18 h in a brain–heart infusion broth (Hopebio, China) supplemented with 5 μg/mL hemin (Sangon Biotech, China) and 0.5 μg/mL vitamin K1 (Sangon Biotech, China). A fresh culture of each strain was collected by centrifugation (5 min at 6000 g) and then washed twice with sterile phosphate-buffered saline (PBS). The final bacterial pellets were resuspended in sterile PBS at a concentration of 5 × 109 colony-forming units (CFUs)/mL.
4
Sensitive Electrochemical Biosensor Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All essential
DNA sequences were synthesized and purified by Shanghai Sangon Biological
Engineering Technology & Services Co., Ltd. (Shanghai, China).
The details of the nucleotide sequences are given inTable S1 . 3,3′,5,5′-Tetramethylbenzidine (TMB)
was purchased from Takara Biotechnology (Dalian, China) Co., Ltd.
Hydrogen peroxide (H2O2), sulfuric acid (H2SO4), and hemin were obtained from Sangon Biotechnology
Co., Ltd. (Shanghai, China). The enzymes, including exonuclease-III
(Exo-III), exonuclease-I (Exo-I), T4 DNA ligase, phi29 DNA polymerase,
and nicking endonuclease (Nb.BbvCI), and the F96 Maxisorpnunc-immuno
plate (96-well plate) were purchased from Thermo Fisher Scientific
Inc. (Shanghai, China). Dimethyl sulfoxide (DMSO) was used to create
a 100 mM hemin stock solution before being further diluted in water.
DNA sequences were synthesized and purified by Shanghai Sangon Biological
Engineering Technology & Services Co., Ltd. (Shanghai, China).
The details of the nucleotide sequences are given in
was purchased from Takara Biotechnology (Dalian, China) Co., Ltd.
Hydrogen peroxide (H2O2), sulfuric acid (H2SO4), and hemin were obtained from Sangon Biotechnology
Co., Ltd. (Shanghai, China). The enzymes, including exonuclease-III
(Exo-III), exonuclease-I (Exo-I), T4 DNA ligase, phi29 DNA polymerase,
and nicking endonuclease (Nb.BbvCI), and the F96 Maxisorpnunc-immuno
plate (96-well plate) were purchased from Thermo Fisher Scientific
Inc. (Shanghai, China). Dimethyl sulfoxide (DMSO) was used to create
a 100 mM hemin stock solution before being further diluted in water.
5
Cultivation and Preservation of Gut Bacteria
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bacteroides ovatus ELH-B2 was recovered from the in-house preservations at Culture Collections of Food Microbiology (CCFM), Jiangnan University (Wuxi, China). B. ovatus JCM5824 was purchased from RIKEN BioResource Center, Japan. Salmonella enterica CMCC50335 and Escherichia coli CMCC44102 were acquired from the National Center for Medical Culture Collections, China.
Salmonella enterica and E. coli were anaerobically cultured in brain heart infusion (BHI, Hopebio, China) at 37°C. The B. ovatus strains were cultured in BHI supplemented with hemin (Sangon Biotech, China) and vitamin K1 (BHIS) at 37°C in anaerobic chamber for further analysis of bacterial characterizations. The bacteria solutions for in vivo tests were prepared with cells at early stationary phase after centrifugation at 6000 rpm for 15 min and re-suspension in phosphate buffer saline supplemented with 20% glycerol, and maintained at -80°C. Cell viability after freezing and thawing was evaluated via colony-forming unit (cfu) enumeration on BHIS agar before use.
Salmonella enterica and E. coli were anaerobically cultured in brain heart infusion (BHI, Hopebio, China) at 37°C. The B. ovatus strains were cultured in BHI supplemented with hemin (Sangon Biotech, China) and vitamin K1 (BHIS) at 37°C in anaerobic chamber for further analysis of bacterial characterizations. The bacteria solutions for in vivo tests were prepared with cells at early stationary phase after centrifugation at 6000 rpm for 15 min and re-suspension in phosphate buffer saline supplemented with 20% glycerol, and maintained at -80°C. Cell viability after freezing and thawing was evaluated via colony-forming unit (cfu) enumeration on BHIS agar before use.
6
Hemolytic Activity Assay of DSM 2950
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The hemolytic activity of DSM 2950 was tested on GAM agar supplemented with 0.01% hemin, vitamin K1, and 5% ovine blood. The hemin and vitamin K1 were purchased from Sangon Biotech (Shanghai, China) Co., Ltd., and sterile defibrated sheep blood was purchased from Bkman Biotechnology (Changde, China) Co., Ltd. First, 100 μL of culture at an appropriate dilution was spread on the agar, which was then anaerobically incubated at 37 °C for 48 h. After incubation, the hemolytic activity was categorized as β-hemolysis (clear halos around colonies), α-hemolysis (greenish halos around the colonies), or γ-hemolysis (the absence of hemolysis). Staphylococcus aureus ATCC29213, which exhibits β hemolysis, was used as a positive control [27 (link)].
7
Hemin-Catalyzed Oxidation of ABTS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The oligonnucleotide strands, Mes buffer, hemin and 2,2′-azinobis(3-ethylbenzothiozoline)-6-sulfonic acid (ABTS) were supplied by Sangon Biotech Co., Ltd. (Shanghai, China). The used DNA sequences were given in follow:
GCDSA: 5′-GGGCTGGGAGGGTTGGGGTATTATTTTTGG-TTGTGCCCTATG-3′; Complementary strands: 5′-CCCAACCCT-3′ (9 bases); 5′-CCCAACCCTCC-3′ (11 bases). DNA concentrations were represented as single-stranded concentrations, which were determined by measuring the absorbance at 260 nm. The final concentrations of hemin and ABTS were 2 μM and 5 mM, respectively, which were stored in freezer far from light. All chemicals in the experiment were analytical grade. Deionized water (18.2 MΩ cm resistivity) was used in this work.
GCDSA: 5′-GGGCTGGGAGGGTTGGGGTATTATTTTTGG-TTGTGCCCTATG-3′; Complementary strands: 5′-CCCAACCCT-3′ (9 bases); 5′-CCCAACCCTCC-3′ (11 bases). DNA concentrations were represented as single-stranded concentrations, which were determined by measuring the absorbance at 260 nm. The final concentrations of hemin and ABTS were 2 μM and 5 mM, respectively, which were stored in freezer far from light. All chemicals in the experiment were analytical grade. Deionized water (18.2 MΩ cm resistivity) was used in this work.
8
Anaerobic Fermentation and Colitis Induction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The standard short-chain fatty acids (SCFAs) solutions, including lactate, acetate, propionate, succinate, and butyrate were all purchased from Sigma-Aldrich (St. Louis, MO, USA). Tryptone, peptone, yeast extract, and Tween 80 used for the in vitro anaerobic fermentation experiments were all obtained from Sigma-Aldrich (St. Louis, MO, USA). Dextran sulfate sodium (DSS) was purchased from MP Biomedicals (Solon, OH, USA). DSS was used to induce ulcerative colitis in the colon of C57BL/6J mice.
Hemin, alginate, agar, and L-cysteine hydrochloride were acquired from Sangon Biotech (Shanghai, China). These chemicals were added to the VI medium and were used for the anaerobic culture of the bacterium B. uniformis F18-22. All other chemicals of analytical grade used in the present study were purchased from Sinopharm Chemical (Shanghai, China) unless otherwise specified.
Hemin, alginate, agar, and L-cysteine hydrochloride were acquired from Sangon Biotech (Shanghai, China). These chemicals were added to the VI medium and were used for the anaerobic culture of the bacterium B. uniformis F18-22. All other chemicals of analytical grade used in the present study were purchased from Sinopharm Chemical (Shanghai, China) unless otherwise specified.
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!