The most commonly used procedures for myofibrillar protein extraction involves denaturing solutions containing urea, thiourea, reducing agents (DTT, beta-mercaptoethanol
), detergents (SDS, sodium dodecyl sulfate), and salts [7, 8].
Presence of beta-mercaptoethanol can increase the glutathione content of pig oocytes matured in vitro and the rate of blastocyst development after in vitro fertilization.
The importance of having high glutathione (GSH) level after bovine in vitro maturation on embryo development: Effect of beta-mercaptoethanol, cysteine and cystine.
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Hoechst 33342 (HO), propidium iodide (PI), sodium pyruvate, glutamine, and beta-mercaptoethanol
were purchased from Sigma (St.
Addition of beta-mercaptoethanol
or trolox at the morula/blastocyst stage improves the quality of bovine blastocysts and prevents induction of apoptosis and degeneration by prooxidant agents.
The cells were grown in RPMI1640 with 10% fetal bovine serum containing 50 [micro]M beta-mercaptoethanol
and 100 units/mL of penicillin and streptomycin.
Then for every 100 mL of BRL conditioned media, we added the following standard cell culture ingredients (generating 150 mL total): KO DMEM (15 mL), FBS (7.5 mL), 100 mM sodium pyruvate (1.5 mL), 100x nonessential amino acids (1.5 mL; Sigma), 100x Penicillin-Streptomycin (1.5 mL, Sigma), 100x nucleosides (1.5 mL, Millipore), 100x Glutamax (250 [micro]L, Life Technology), and beta-mercaptoethanol
(190 [micro]L, Sigma) to generate "BRL conditioned plus" media, with the final concentrations as listed in Table 1.
The samples were mixed with 1x sample buffer to have finally 0.06 M Tris, pH 6.7, 2% SDS, 10% glycerol, 0.001 bromophenol blue, and 0.1% by volume beta-mercaptoethanol
just before loading.
Promoting effect of beta-mercaptoethanol
on in vitro development under oxidative stress and cystine uptake of bovine embryos.
In a separate experiment, a 60-fold molar excess of beta-mercaptoethanol
(B-ME) was added to Cu[Cl.sub.2]-treated hemoglobin, incubated for 1 h, and dialyzed for 3 h before G-100 chromatography.
MIN6 cells were cultured in DMEM supplemented with 15% FBS, 100 U/mL penicillin, 100 [micro]g/mL streptomycin, 2mM L-glutamine, and 70 [micro]mol/L beta-mercaptoethanol
. Cells were divided into NC, A, B, and C groups at 6 x [10.sup.5]/mL; medium was changed every two days, and A, B, and C groups were treated with 2, 5, and 10 [micro]M simvastatin, respectively, for 48 h.
enhances blastocyst formation rate of bovine in vitro-matured/in vitro-fertilized embryos.