For reducing SDS-PAGE, 80 [micro]L of sample (2 mg/mL) was mixed with 20 [micro]L of 10% SDS, 2 [micro]L of 2-mercaptoethanol mercaptoethanol
and 1 [micro]L of 1% w/v bromophenol blue.
The presence of 50 kDa mTcTXNPx and 46 kDa cTcTXNPx, both in the oxidized form, (8) is not observed in samples treated with mercaptoethanol
, (9) which likely prevents the formation of disulphide bonds and therefore prevents dimerization.
Then 20 ug protein were mixed with SDS-PAGE sample buffer and mercaptoethanol
to incubation at 100degC for 5 minutes, separated by 15% SDS-PAGE and transferred to PVDF membrane.
The effect of surfactants and inhibitors including mercaptoethanol
, EDTA (1 mM), SDS (1%, w/v), Tween (20 and 80; 0.1%, v/v), and Triton X-100 (0.1%, v/v) on pectinase enzyme activity was studied by directly incorporating them into the enzyme substrate system.
The most commonly used approaches to induce differentiation of MSCs into neuron-like cell include (1) antioxidant agents such as mercaptoethanol
, dimethylsulfoxide (DMSO), and beta hydroxy acid (BHA); (2) neurotropic factors such as retinoic acid (RA), nerve growth factor (NGF), epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), brain derived neurotropic factor (BDNF); (3) Chinese herbs and their effective components such as astragalus glycosides, berberine, tanshinone, lycium barbarum polysaccharide, acanthopanax; (4) cultural method; and (5) gene transfection: transfection of specific genes such as Noggin  and Notch into hUC-MSCs to get neuron-like cell.
Under the reducing action of P mercaptoethanol
the homogenous 170 kDa band disappeared and few narrow bands with lower molecular weight were visible.
Then enzyme was chromatographed in a Sephadex G-75 column (4.5x 85 cm) that was equilibrated and eluted with 0.01M acetate buffer pH 5.1, containing 0.1M NaCl, 1mM EDTA, 1mM PMSF and 1mM AY- mercaptoethanol
. The active fractions were combined and the enzyme was concentrated by Amicon ultrafiltration using Ym 5 membrane to 7 mL.
Yeast cells were harvested by centrifugation at 4000 rpm for 10 min and an appropriate amount of lysis buffer (SDS and mercaptoethanol
: 1:1 Check spelling) was added to the cell pellet and then sonicated at 4oC for 15 s with a 60 s interval 2-3 times and centrifuged at 4000 rpm for 10 min.
USP solution consists of 4 M guanidinium hydrochloride (GuHCL) (Amresco), 50mM Tris-CI, (Sigma), 25 mM EDTA (SRL), 0.5% Sarkosyl (Sigma) and 0.2 M mercaptoethanol
HCl, petroleum ether, CaCO3 were purchased from Fluka Chemicals and chloroform, tris, KCl, mercaptoethanol
were purchased from Merck.
The antioxidants supplementation to the culture medium can be divided into two large groups: ones that are non-enzymatic or metabolic with low molecular weight, such as L-ascorbic acid (vitamin C), cysteine, alpha-tocopherol, and Beta- mercaptoethanol
and ones with an enzymatic action, such as superoxide dismutase, glutathione peroxidase and catalase (Nordberg and Arner,
In 1971, it was firstly reported that OPA could react with amino acid and ammonium in the existence of mercaptoethanol
to produce a strongly fluorescent compound .