Organics Background

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TCEP-HCl
OmniPur® TCEP-HCl– Water-soluble, odorless • More stable and effective than DTT • Acts over a broad pH range (pH 1.5 to 8.5) • Fast acting • More resistant to air-oxidation than DTT

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BMS has been shown to reduce accessible disulfide bonds 5 to 7 times faster than DTT.


BMS [Bis (2-mercaptoethyl)sulfone]- Under nondenaturing conditions at pH 7, in certain samples, BMS has been shown to reduce accessible disulfide bonds 5 to 7 times faster than DTT. Water-soluble • Low pKa • Stable in solution • High reduction potential • Acts faster than DTT • Works even on less accessible disulfide bonds

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Below are highlights and summaries from Essential Biochemicals for Research, which is a technical resource and product guide for scientists for the preparation and use of biochemicals.To view the full content, please download or request a print copy of this resource.

Albumins

Albumin is a ~66 kDa serum protein and the most abundant in mammalian blood plasma. It helps maintain osmotic pressure and serves as a carrier protein, delivering hormones, nutrients, fatty acids and other molecules to cells. Due to its several intramolecular disulfide bonds, albumin has high structural stability, enabling it to resist denaturation under many conditions.

Amino Acids

Peptides and polypeptides are polymers of α-amino acids. There are 20 α-amino acids that make up all proteins of biological interest. The α-amino acids in peptides and proteins consist of a carboxylic acid (-COOH) and an amino (-NH2) functional group attached to the same tetrahedral carbon atom. This carbon is known as the α-carbon. The type of R-group attached to this carbon distinguishes one amino acid from another.

IPTG (Isopropyl-β-D-thiogalactopyranoside)

Expression of the three genes in the lac operon of E. coli is under the negative control of a repressor protein consisting of four identical 38.6 kDa subunits. IPTG, a free-flowing, non-hygroscopic carbohydrate, induces expression of the lac operon by allosterically interacting with the repressor molecule and causing a 300-fold reduction in the repressor’s affinity for the operator sequence. The action of IPTG leads to the production of the lac operon gene products, which allows E. colito utilize lactose as an energy source.

Commonly Used Reducing Agents

Cleland’s Reagent or Dithiothreitol (DTT)
Dithiothreitol (DTT), also known as Cleland’s Reagent, is a water-soluble, protective reagent for sulfhydryl groups and is widely used for the study of disulfide exchange reactions of protein disulfides.

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TCEP, Hydrochloride

TCEP is considered to be a more potent, stable and effective alternative to DTT. It can reduce disulfide bonds even in strongly acidic environments (pH 1.5 to 4.5) where DTT is essentially ineffective.

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BMS [Bis (2-mercaptoethyl)sulfone]
BMS is a water-soluble reagent useful for the reduction of native disulfide bonds in proteins, and is considered to be a superior reducing agent to DTT.

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