Technical Data
T5010-10
Thioredoxin (fusion protein)
Description:
Thioredoxin catalyzes disulfide exchange in proteins via an oxidation/reduction reaction mechanism and is used for apoptosis inhibition in human PBL blasts and Jurkat T-cells. Thioredoxin also functions as a catalyst for filamentous phage assembly and as an accessory protein for phage T7 DNA Polymerase.

Recombinant DNA technology allows the addition of short pieces of well-defined tags, "peptides" or proteins at the amino or c-terminus of target genes, which can provide 'affinity handles' designed to bind specific matrices. Therefore, tags enable a selective identification and purification of the protein of interest. Eukaryotic genes are often cloned into E. coli thioredoxin gene, resulting in the expression of a desired protein as a fusion hybrid with thioredoxin (1). Thioredoxin is a small electron transport protein that serves as hydrogen door in the enzymatic reduction of ribonucleotides to deoxyribonucleotides (1). Thioredoxin from E. coli consists of a single polypeptide of 108 aa (~11.7kD). The gene encoding Thioredoxin has been clones into several plasmids and serves as a fusion partner for the expression of other clones genes. The system often produces high expression of soluble fusion proteins in the E. coli cytoplasm. The fusion proteins may also fold correctly with biological activity.

Anti-Thioredoxin antibodies allow a simple isolation of fusion proteins directly from the crude bacterial lysate, using immunoaffinity chromatography (1) or used in immunoprecipitation. Anti- thioredoxin can also be used for the immunocytochemical detection of thioredoxin in cells and tissues that express transfected bacterial thioredoxin gene (1). Ant-thioredoxin may be used in various immunoassays to identify the expression of thioredoxin fusion protein.

Applications:
Suitable for use in ELISA and Western Blot. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000-1:5000 using ECL. Antibodies react with native and denatured THRx-tag containing proteins.
ELISA: 1:10-50K using 50-100ng control antigen/well).
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
PabIgGSerum
SizeStorageShippingSourceHost
100ul-20CBlue IceRabbit
Concentration:
As reported
Immunogen:
Recombinant purified E. coli thioredoxin.
Purity:
Neat serum
Form
Supplied as a lyophilized powder from PBS, 0.05% sodium azide.
Specificity:
Recognizes rabbit Thioredoxin.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Narayanan, S., J. Chromatogr., 658, 237 (1994).
2. Casey, J., et al., J. Immunol. Meth., 179, 105 (1995).
3. Uhlen, M., and Moks, T., Meth. Enzymol., 185, 129 (1990).
4. Skerra, A., et al., Bio/Technology, 9, 273 (1991).
5. Holmgren A (1985) Ann Rev. Biochem. 54, 237; Rosen A et al (1995) Int. Immunol. 7, 625