Certificate of Analysis and Data Sheet

Source:
E.Coli

Catalog No.
ENZ-276

Background:

DsbA appears to be necessary for correct formulation of disulfide bonds in exported proteins in vivo. DsbA is useful as a standard in immunoblotting. This protein catalyses the reduction and exchange of disulfide bonds and the oxidation of free sulfhydryl groups in vitro. It is the strongest oxidant of the thioredoxin superfamily. This thio/disulfide oxidoreductase is required for efficient disulfide bond formation in the periplasm of E. coli. dsbA is purified by proprietary chromatographic techniques.

Description :

DsbA produced in E.Coli is a periplasmic protein isolated from E. coli having a molecular mass of 23,149 Dalton dsbA is purified by proprietary chromatographic techniques.

Physical Appearance:

Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation:

dsbA was lyophilized after from a sterile solution containing 50mM sodium phosphate buffer and 100mM sodium chloride..

Solubility:

It is recommended to reconstitute the lyophilized Human dsbA in sterile 18MO-cm H2O not less than 100ug/ml, which can then be further diluted to other aqueous solutions.

Stability:

Lyophilized dsbA although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution dsbA should be stored at 4 C between 2-7 days and for future use below -18 C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.

Purity:

Greater than 95.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.

Amino acid sequence:

The sequence of the first five N-terminal amino acids was determined and was found to be Met-Ly-Lys-Ala-Trp.

Dimers and aggregates:

Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

Applications:

1.Western blot
(we recommend that the material be diluted in 1X SDS-PAGE sample buffer (1). On a 15-well minigel system, 50ng of protein per lane should be sufficient when used in a colorimetric Western Blot with D9625 at a dilution of 1:10,000 as the primary antibody and an appropriate alkaline phosphatase conjugated secondary antibody for detection.)

Protein content:

Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm.
2. Analysis by RP-HPLC, using a calibrated solution of dsbA as a Reference Standard.

Endotoxin:

Less than 0.1 ng/ug (IEU/ug) of dsbA .

Usage:

This material is offered by ProSpec-TechnoGene for research, laboratory or further evaluation purposes.

Protein Family:

Protein Domains:

Domains:
•IPR006662   Thioredoxin-related
•IPR001853   DSBA oxidoreductase

Protein links:

Precursor- Protein structure and amino acid sequence:

Latest Publications:

1. Mechanistic implications of the structure of the mixed-disulfide intermediate of the disulfide oxidoreductase, 2-ketopropyl-coenzyme m oxidoreductase/carboxylase(,).
Biochemistry 2006 Jan 10;45(1):113-20
2. Insights on a New PDI-like Family: Structural and Functional Analysis of a Protein Disulfide Oxidoreductase from the Bacterium Aquifex aeolicus.
J Mol Biol 2006 Feb 10;356(1):155-64
3. Temperature-, SDS-, and pH-induced conformational changes in protein disulfide oxidoreductase from the archaeon Pyrococcus furiosus: a dynamic simulation and fourier transform infrared spectroscopic study.
J Proteome Res 2005 Nov-Dec;4(6):1972-80
4. Characteristics of thiol:protein disulfide oxidoreductase from wheat (Triticum aestivum L.) grain.
Biochemistry (Mosc) 2005 Aug;70(8):935-40
5. The thiol-disulfide oxidoreductase system in the cold-adapted bacterium Pseudoalteromonas haloplanktis TAC 125: discovery of a novel disulfide oxidoreductase enzyme.
Extremophiles 2005 Sep 23;
6. Crystallization and preliminary X-ray diffraction studies of a protein disulfide oxidoreductase from Aquifex aeolicus.
Acta Crystallogr D Biol Crystallogr 2004 Nov;60(Pt 11):2076-7

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中華民國95年06月06日更新