D pantothenic acid
Manufactured by Merck Group
Sourced in Germany
D-pantothenic acid is a water-soluble vitamin that plays a crucial role in various metabolic processes. It is an essential component of coenzyme A, which is involved in the metabolism of fats, carbohydrates, and proteins. D-pantothenic acid is widely distributed in nature and can be found in a variety of food sources, such as meat, dairy products, legumes, and whole grains.
Automatically generated - may contain errors
Lab products found in correlation
Kanamycin, by Merck Group (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Biotin, by Merck Group (2 mentions)
Sypro orange dye, by Thermo Fisher Scientific (1 mentions)
D 1 14c pantothenate, by American Radiolabeled Chemicals (1 mentions)
3h atp, by PerkinElmer (1 mentions)
Ni nta resin, by Qiagen (1 mentions)
Tyloxapol, by Merck Group (1 mentions)
L leucine, by Merck Group (1 mentions)
Casamino acid, by BD (1 mentions)
Hmsc adipogenic differentiation medium bulletkit, by Lonza (1 mentions)
Hygromycin, by Roche (1 mentions)
Streptomycin, by Merck Group (1 mentions)
L lysine, by Merck Group (1 mentions)
7 protocols using d pantothenic acid
1
Radiolabeled Pantothenate Binding Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
d -[1-14C]Pantothenate (specific activity, 55 mCi/mmol) from American Radiolabeled Chemicals; Ni-NTA resin from Qiagen; [3H]ATP from Perkin Elmer (specific activity, 29.8 Ci/mmol); Sypro Orange dye from Thermo Fisher Scientific; d -[3H]pantothenate (specific activity, 50 Ci/mmol) from American Radiolabeled Chemicals; d -pantothenic acid, hemicalcium salt from Sigma-Aldrich; pantothenate-free DMEM from Thermo Fisher Scientific; CoA thioesters from Avanti Polar Lipids. All other materials were reagent grade or better.
+ Expand
2
Mycobacterium Cultivation Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Stock cultures of M. tuberculosis H37Rv ATCC 27294 and M. smegmatis mc2155 were stored frozen at -80°C in Proskauer-Beck media (50% v/v glycerol) and glycerol stock media (50% v/v glycerol, 7H9, ADC, Tween 80), respectively. For propagation of initial culture, frozen stocks were thawed and sub-cultured in Middlebrook 7H9 media with OADC (0.005% oleic acid, 0.5% bovine serum albumin fraction V, 0.2% dextrose, 0.0003% catalase), 0.2% glycerol, and 0.05% Tween 80 until reaching a desirable optical density (OD) for each experiment. 7H9 and OADC were purchased from BD (Franklin Lakes, NJ, USA). Glycerol and Tween 80 were purchased from Sigma-Aldrich (St. Louis, MO, USA). All experiments using virulent M. tuberculosis H37Rv were done in a BSL3 laboratory located at Colorado State University. For experiments using the BSL2 strain, M. tuberculosis H37Rv mc2 6206, bacteria was grown in 7H9 media supplemented with OADC, 0.2% casamino acid (BD, USA), 0.05% tyloxapol, 0.005% L-leucine, 0.0048% D-pantothenic acid, and 0.0025% kanamycin (Sigma-Aldrich, USA). M. tuberculosis H37Rv mc2 6206 was a kind gift from Dr. William R. Jacobs Jr. at Albert Einstein College of Medicine [34 (link)].
3
SGBS Adipocyte Differentiation and Treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human Simpson-Golabi-Behmel syndrome (SGBS) cells were used as a model system of adipocyte biology20 (link). Cells were passaged in DMEM/F-12 (1:1), 100 U/ml penicillin, 100 μg/ml streptomycin (Life Technologies, Darmstadt, Germany), 17 μM D-pantothenic acid and 33 μM biotin (Sigma-Aldrich, Munich, Germany) (referred to as basal medium) supplemented with 10% fetal bovine serum. For the induction of adipogenic differentiation, subconfluent cell cultures were washed with PBS and adipogenic medium (serum-free basal medium with 20 nM human recombinant insulin, 100 nM cortisol, 200 pM triiodothyronine and 10 μg/ml transferrin) supplemented with 2 μM rosiglitazone, 25 nM dexamethasone and 250 μM isobutylmethylxanthine was added. After four days, the medium was changed to adipogenic basal medium alone. On day 14 of adipogenesis TRAIL or an equal amount of vehicle (PBS + 0.01% bovine serum albumin) was added directly to the media.
4
Heterologous Expression of Bmp1 from M. mediterranea
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
As shown in Supplementary Fig. 6 , M. mediterranea MMB-1 bmp1 was ligated into RSFDuet-1 vector MCS-1 digested with BamHI and HindIII restriction enzymes. This construct encodes a N-His6 tag fused to Bmp1. In MCS-2 of the same vector, the ORF corresponding to an ACP synthase (GenBank: NC_015276.1) was inserted using NdeI and XhoI restriction sites. RSFDuet-1 vector utilizes a kanamycin resistance marker. M. mediterranea MMB-1 bmp3 was ligated into CDFDuet-1 vector MCS-2 digested with NdeI and XhoI restriction enzymes. CDFDuet-1 vector utilizes a streptomycin resistance marker. Also shown in Supplementary Fig. 6 is the di-domain architecture of Bmp1. As identified by sequence homology, the N-terminal 77 residues constitute the ACP domain, with Ser35 being the site for phosphopantetheinylation. Residues 78 to the C terminus constitute the TE domain, with Ser202 being the catalytic nucleophilic residue of a typical serine protease/esterase Ser-His-Asp/Glu triad. To generate Bmp1(ACP), the codon corresponding to residue 78 of bmp1 was mutated to a stop codon by site directed mutagenesis. The two vectors were then co-transformed into E. coli BL21Gold(DE3) competent cells. Protein expression cultures were spiked with 0.1 mM D-pantothenic acid (Sigma-Aldrich, P2250-5G), and 10 mg riboflavin. Bmp1(ACP) was then purified by affinity chromatography.
5
Adipogenic Differentiation of SGBS, ASC, and BMSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SGBS cells and ASC were cultured in basal medium (DMEM/F-12 (1:1), 100 U/mL penicillin, 100 µg/mL streptomycin (Life Technologies, Darmstadt, Germany), 17 µM D-pantothenic acid, and 33 µM biotin (Sigma-Aldrich, Munich, Germany)) supplemented with 10% fetal bovine serum.
Subconfluent SGBS cells and ASCs were induced to undergo adipogenic differentiation under serum-free conditions in basal medium supplemented with 20 nM human recombinant insulin, 100 nM cortisol, 200 pM triiodothyronine, 10 µg/mL transferrin, 2 μM rosiglitazone, 25 nM dexamethasone, and 250 µM isobutylmethylxanthine. 4 days after induction, the medium was renewed leaving out rosiglitazone, dexamethasone, and isobutylmethylxanthine. Cells were analyzed on day 14 of differentiation. Adipogenic differentiation of BMSCs was performed using the hMSC Adipogenic Differentiation Medium BulletKit (Lonza, Basel, Switzerland) according to the manufacturer’s instruction.
Adipogenic differentiation rate was determined by counting lipid filled adipocytes with at least 5 visible lipid droplets and pre-adipocytes using a net micrometer, followed by dividing the number adipocytes by the total cell number.
Subconfluent SGBS cells and ASCs were induced to undergo adipogenic differentiation under serum-free conditions in basal medium supplemented with 20 nM human recombinant insulin, 100 nM cortisol, 200 pM triiodothyronine, 10 µg/mL transferrin, 2 μM rosiglitazone, 25 nM dexamethasone, and 250 µM isobutylmethylxanthine. 4 days after induction, the medium was renewed leaving out rosiglitazone, dexamethasone, and isobutylmethylxanthine. Cells were analyzed on day 14 of differentiation. Adipogenic differentiation of BMSCs was performed using the hMSC Adipogenic Differentiation Medium BulletKit (Lonza, Basel, Switzerland) according to the manufacturer’s instruction.
Adipogenic differentiation rate was determined by counting lipid filled adipocytes with at least 5 visible lipid droplets and pre-adipocytes using a net micrometer, followed by dividing the number adipocytes by the total cell number.
6
Mycobacterial Strain Cultivation Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All strains included in this study are listed in Table S2 in the supplemental material. Mycobacterial strains were routinely grown on Middlebrook 7H10 agar with 10% oleic acid-albumin-dextrose-catalase (OADC; Difco) or in Middlebrook 7H9 liquid medium with 10% ADC (Difco) and 0.05% Tween 80. Additional supplement was necessary for culturing of M. tuberculosis mc26020 (ΔlysA ΔpanCD), for which 100 ng/mL l -lysine (Sigma) and 25 ng/mL d -pantothenic acid (Sigma) were included. Appropriate antibiotics were added when necessary, i.e., 50 μg/mL kanamycin (Sigma), 50 μg/mL hygromycin (Roche), or 30 μg/mL streptomycin (Sigma). M. marinum and M. tuberculosis cultures and plates were incubated at 30°C and 37°C, respectively. Escherichia coli DH5α was used for cloning and was grown on LB agar plates at 37°C. Antibiotics were added at the same concentrations as described above.
7
Heterologous Expression of Bmp1 from M. mediterranea
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!