Bifidobacterium longum subsp. longum H3-R1, H9-5, H10-1, H16-2, H20-14, and H27-10, Bifidobacterium animalis subsp. lactis H15-2, H27-9, and H34-21, Bifidobacterium bifidum H3-R2 and H10-5, B. longum subsp. infantis H5-21 and H11, Bifidobacterium breve H34-14 and Bifidobacterium pseudocatenulatum H17-2 were isolated from exclusively breast-fed infants and stored at the Key Laboratory of Dairy Science (KLDS), Ministry of Education, China. B. animalis subsp. lactis BB12 (ATCC 27536) was used as a reference strain for screening bifidobacteria with immunostimulatory activity. All strains were anaerobically incubated in De Man, Rogosa and Sharpe (MRS) (Oxoid, United Kingdom) medium supplemented with 0.05% L-cysteine hydrochloride (mMRS) at 37°C for 18 h and were sub-cultured twice prior to the experiment. Bacteria were harvested by centrifugation (8000 rpm for 5 min at 4°C) and the cells were washed with saline buffer. The cell pellets were resuspended to the dosages as specified in the relevant sections for the in vitro and in vivo work.
L cysteine hydrochloride
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
L-cysteine hydrochloride is a chemical compound commonly used in laboratory settings. It serves as a source of the amino acid cysteine. The product is available in various purities and formulations to meet the needs of different applications.
Automatically generated - may contain errors
Lab products found in correlation
α ketoglutarate, by Thermo Fisher Scientific (1 mentions)
Horse blood, by Thermo Fisher Scientific (1 mentions)
Anaerogen gas pack, by Thermo Fisher Scientific (1 mentions)
Tryptic soy agar, by Thermo Fisher Scientific (1 mentions)
Haemin, by Thermo Fisher Scientific (1 mentions)
Nitric acid, by Thermo Fisher Scientific (1 mentions)
Hydrochloric acid (hcl), by Thermo Fisher Scientific (1 mentions)
Celltiter 96 aqueous one solution cell proliferation assay, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Quant it picogreen dsdna assay kit, by Thermo Fisher Scientific (1 mentions)
Exendin 4 ex 4, by AnaSpec (1 mentions)
Iscript cdna synthesis kit, by Bio-Rad (1 mentions)
Itaq universal sybr green supermix, by Bio-Rad (1 mentions)
Rneasy plus mini kit, by Qiagen (1 mentions)
Qiazol lysis reagent, by Qiagen (1 mentions)
Insulin, by ProSpec (1 mentions)
Anaerogen, by Thermo Fisher Scientific (1 mentions)
Nutrient broth, by Thermo Fisher Scientific (1 mentions)
Ellman s reagent, by Thermo Fisher Scientific (1 mentions)
Cytation 5, by Agilent Technologies (1 mentions)
9 protocols using l cysteine hydrochloride
1
Immunostimulatory Bifidobacterium Strains
2
Cultivation and Isolation of L. pneumophila
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
L. pneumophila wild-type strain Heysham-1 (ATCC 43107), the mutant-type strain (T6-K13), and the complementant strain (T6-K24-1) were cultured at 37 °C for 3 days on ACES-buffered charcoal-yeast extract (BCYE) agar containing 0.4 mg/mL L-cysteine hydrochloride, 0.25 mg/mL ferric pyrophosphate, and 0.5 mg/mL α-ketoglutarate (Oxoid, Hampshire, UK) [34 (link)]. The bacterial mass collected from plates was suspended in 0.5 M NaCl and centrifuged at 8000 rpm for 20 min. The bacterial pellet was washed once with 0.5 M NaCl and once with MQ water. Then, 5 g of lyophilized bacterial mass was used to isolate lipopolysaccharide and 130 mg of lyophilized bacterial mass was used to isolate lipids. The yield of LPS was 1.5% for the Heysham-1 strain and T6-K24-1, and 1.2% for the mutant-type strain. Lipids accounted for 6.8% of the Heysham-1 biomass, 6.6% of the T6-K13 strain, and 7.3% of the complementant strain T6-K24-1.
L. pneumophila strains were cultured in N-(2-acetamido)−2-aminoethanesulfonic acid (ACES)-buffered yeast extract broth (BYE) at 37 °C for 24 h. Liquid cultures were used for adhesion, invasion, and infection tests.
L. pneumophila strains were cultured in N-(2-acetamido)−2-aminoethanesulfonic acid (ACES)-buffered yeast extract broth (BYE) at 37 °C for 24 h. Liquid cultures were used for adhesion, invasion, and infection tests.
3
Culturing and Manipulating P. gingivalis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The P. gingivalis strains used in this study are outlined in Table S1. All growth media and supplements were obtained from Sigma-Aldrich (Co. Wicklow, Ireland) unless stated otherwise. P. gingivalis strains were grown on Tryptic Soy Agar (TSA) supplemented with 5 g/l yeast extract, 0.50 g/l L-cysteine hydrochloride, 5.0 µg/ml haemin and 1.0 µg/ml menadione and 5% horse blood (Oxoid Limited, Basingstoke, UK). Bacteria were incubated at 37°C in BBL gas jars under anaerobic conditions generated by AnaeroGen gas packs (Oxoid). When required media was supplemented with 5.0 µg/ml erythromycin, 1.0 µg/ml tetracycline or 10 µg/ml chloramphenicol. For liquid cultures supplemented Tryptic Soy Broth (sTSB) containing identical supplements was used. Plasmid pJW1 was obtained from Prof. A. Progulske-Fox, University of Florida (Lépine et al., 1996) . The ATCC33277 fimA mutant was obtained from Hua Xie, School of Dentistry, Meharry Medical College, Nashville TN (Zheng et al., 2011) .
4
Synthesis of Gold Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Betulin, nitric acid, hydrochloric acid and L -Cysteine hydrochloride were purchased from Fisher Scientific (Loughborough, United Kingdom). Chloroauric acid (HAuCl4), trisodium citrate dihydrate (C6H5O7Na3⋅2H2O), and poly(ethylene glycol) 2-mercaptoethyl ether acetic acid (HOOC-Peg-SH, MW 3500) were purchased from Sigma-Aldrich (Taufkirchen, Germany) and used as received. The deionized water was prepared by using the Ultrapure, Millipore® Direct-Q® 3 with UV radiation.
5
Halloysite Nanotubes for Stem Cell Delivery
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Poly(caprolactone) (PCL; Mw-80kD), cellulose acetate (CA; Mw-50kD), halloysite nanotubes (HNTs), fluorescein isothiocyanate (FITC), papain, other salts, and analytical grade solvents were purchased from Sigma-Aldrich (St. Louis, MO). Human mesenchymal stem cells were purchased from RoosterBio Inc. (Frederick, MD) and Dulbecco's minimum essential medium (DMEM) media from Lonza Bioscience (Morrisville, NC). All cell culture supplies including pen-strep (P/S), fetal bovine serum (FBS), dialysis bags, l -cysteine hydrochloride, 0.25% trypsin EDTA, Quant-iT™ PicoGreen™ dsDNA Assay Kit, and Cell Titer 96® AQueous One Solution Cell Proliferation Assay were purchased from Fisher Scientific (Fair Lawn, NJ). QIAzol Lysis Reagent and RNeasy Plus Mini kit were purchased from QIAGEN Inc. (Germantown, MD) while iScript cDNA Synthesis Kit and iTaq Universal SYBR Green Supermix were purchased from Bio-Rad (Hercules, CA). Exendin-4 (Ex-4) was purchased from AnaSpec Inc. (Fremont, CA), and insulin was purchased from ProSpec Inc. (East Brunswick, NJ).
6
Cryopreservation of Probiotic and Pathogenic Bacteria
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
B. bifidum ATCC 11863 and the commercial probiotic strains, L. acidophilus LA-5 ® and B. lactis BB-12 ® were grown respectively in Reinforced Clostridial Medium (RCM; Oxoid, Basingstoke, UK) and de Man Rogosa Sharpe broth (MRS, Oxoid) supplemented with 0.05% (w/v) L-cysteine hydrochloride (Fisher Scientific, UK) "MRSc" under anaerobic conditions (anaerobic jar with AnaeroGen, Oxoid to generate the anaerobic environment) at 37 o C for 48 h . P. aeruginosa was grown in Nutrient broth (NB; Oxoid) aerobically at 37 o C. The cells were harvested when they reached stationary phase of growth. The cells were washed in phosphate buffered saline (PBS), and resuspended in 15% (v/v) glycerol at an organism density of 10 8 CFU/mL and frozen in 1.8 mL aliquots over liquid nitrogen (Beezer et al. 1976) .
Aliquots were stored under liquid nitrogen until required.
Aliquots were stored under liquid nitrogen until required.
7
Quantification of Free Sulfhydryl Groups
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The free sulfhydryl group content of the samples was analyzed following the method of Beveridge et al. [31 (link)] and manufacturer’s instructions with some modifications. 5,5′-Dithio-bis-(2-Nitrobenzoic Acid) (DTNB) and Ellman’s reagent (Thermo Fisher Scientific, Waltham, MA, USA) were utilized to determine the content of the free sulfhydryl group in the samples. Sodium phosphate buffer (0.1 M, pH 8.0) was used to dilute the protein solution to a certain concentration. l -Cysteine hydrochloride (Alfa Aesar, Tewksbury, MA, USA) was used as a standard. Dilutions of cysteine (0.25–1.5 mM) sodium phosphate buffer (0.1 M, pH 8.0) were prepared to plot a standard curve. An amount of 50 μL of Ellman’s reagent solution (4 mg/mL) was added in the mixture of 250 μL of protein sample and 2.5 mL of sodium phosphate buffer. After incubation at room temperature for 15 min, the absorbance was measured in triplicates at 412 nm using a multimode microplate reader (Cytation 5, BioTek Instruments, Inc., Winooski, VT, USA). The free sulfhydryl group content was expressed as μmol/g protein.
8
Hydrogel Synthesis and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Poly(acrylic
acid) (PAA) (450 kDA), acrylic
acid (AA), and 2-hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophhenone
(Irgacure 2959) were purchased from Sigma-Aldrich (Ireland).l -Cysteine hydrochloride was purchased from Alfa Aesar (Ireland).
1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was purchased
from Tokyo Chemicals (United Kingdom). All other chemicals used were
analytical grade and were used without further purification.
acid) (PAA) (450 kDA), acrylic
acid (AA), and 2-hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophhenone
(Irgacure 2959) were purchased from Sigma-Aldrich (Ireland).
1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was purchased
from Tokyo Chemicals (United Kingdom). All other chemicals used were
analytical grade and were used without further purification.
9
Synthesis of Metal Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HAuCl4 ∙ 3H2O (99.99% (metals basis)), L-cysteine hydrochloride (98%), D-cysteine hydrochloride (98%), L-glutathione (98%+), D-methionine (99%), L-methionine (98 + %), L-cystine dihydrochloride (98%), D-cystine (98%), Cu(NO3)2 trihydrate (98%), Fe(NO3)3 nonahydrate (98%), Co(NO3)2 hexahydrate (99%), Ni(NO3)2 hexahydrate (98%), and KOH (99 + %) were purchased from Alfa Aesar. AgNO3 (>99.9%), HCl (37%), and H2SO4 (75%) were purchased from Roth. Hexadecyl-trimethylammonium bromide (CTAB, >99%), ascorbic acid (AA, >99%), and acetic acid (80%), were purchased from Acros Organics. Hexadecyltrimetryammonium chloride (CTAC, >98.0%), 1-decanol (>98%), L-penicillamine (99%), D-penicillamine (99%), Cd(NO)3 tetrahydrate (99%), Pb(NO3)2 (99%), and ammonium hydroxide (28%–30%) were purchased from Sigma-Aldrich. Milli Q water was used in all experiments. All reagents were used as received without further purification.
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!