Recombinant Human Thioredoxin-1 Protein, CF

Catalog # Availability Size / Price Qty
1970-TX-500
Product Details
Citations (4)
FAQs
Reviews

Recombinant Human Thioredoxin-1 Protein, CF Summary

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to catalyze the reduction of insulin. The reaction leads to precipitation, which can be measured by absorbance at 650 nm. The specific activity is >10 A650/min/mg, as measured under the described conditions.
Source
E. coli-derived human Thioredoxin-1 protein
Val2-Val105
Accession #
N-terminal Sequence
Analysis
Val2
Predicted Molecular Mass
12 kDa
SDS-PAGE
12 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

1970-TX

Formulation Lyophilized from a 0.2 μm filtered solution in PBS, EDTA and DTT.
Reconstitution Reconstitute at 1000 μg/mL in sterile PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Assay Procedure

Materials
  • Assay Buffer: 50 mM MES, 250 mM NaCl, 2 mM EDTA, pH 6.5
  • Recombinant Human Thioredoxin‑1 (rhTRX-1) (Catalog # 1970-TX)
  • Human Insulin (Sigma, Catalog # I-9278)
  • Dithiothreitol (DTT) (Sigma, Catalog # D-0632), 1 M stock in deionized water
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: Spectramax Plus by Molecular Devices) or equivalent
  1. Dilute rhTRX-1 to 20 µM in Assay Buffer.
  2. Dilute hInsulin to 520 µM in Assay Buffer.
  3. Dilute DTT to 55 mM in Assay Buffer.
  4. Load into a clear plate 50 µL of Assay Buffer, 25 µL of 20 µM rhTRX-1, and 25 µL of 520 µM hInsulin. For a Substrate Blank, load 75 µL of Assay Buffer and 25 µL of 520 μM hInsulin. Start the reaction by adding 10 µL of 55 mM DTT to all wells.
  5. Read at Abs650 in kinetic mode for 15 minutes.  Use the linear portion of the reaction to determine specific activity.
  6. Calculate specific activity:

     Specific Activity (Abs/min/mg) =

Adjusted Vmax* (Abs/min)**
amount of enzyme (mg)

     *Adjusted for Substrate Blank
     **Note: the output of many spectrophotometers in kinetic mode is in mOD

Per Well:
  • rhTRX-1: 0.00585 mg
  • hInsulin: 118 µM
  • DTT:  5 mM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Thioredoxin-1

Thioredoxins (Trxs) are a group of small ubiquitous proteins in all living cells that are key regulators of cellular redox balance (1, 2). The mammalian Trx family has three members. The Trx-1, which is a secreted and cellular protein, the mitochondria-specific Trx-2, and the Trx-like cytosolic protein p32TrxL (3‑5). The active site of mammalian Trxs contains two cysteines in the conserved sequence -Y-C-G-P-C-K-. In Trx-1 the conserved cysteine residues are in positions 32 and 35, respectively. Trxs exist either in a reduced or in an oxidized state when the two cysteines at the active site form an intramolecular disulfide bridge. NADPH and the flavoprotein thioredoxin reductase can convert the oxidized Trx into the reduced Trx. Trx-1 is the only extracellular occurring thioredoxin, and is secreted by lymphocytes, hepatocytes, fibroblasts, and several tumor cells. Plasma concentrations of Trx-1 are up to 6 nM (6). In cells, Trx-1 is localized predominantly in the cytoplasm. Small amounts have been detected in the nucleus and in association with the outside surface of the cells. Expression of Trx-1 is increased under various stress conditions such as hypoxia, elevated hydrogen peroxide concentrations, photochemical oxidative stress, and viral and bacterial infections. Biological functions of Trx-1 include growth factor activity, antioxidant properties, a cofactor that provides reducing equivalents, and transcriptional regulation (1, 2). The synovial tissue of rheumatoid arthritis patients produces increased levels of Trx-1 under oxidative stress conditions, and a correlation exists between the plasma levels of Trx-1 and the severity of the disease, making Trx-1 a biomarker for this pathological condition (7, 8).

References
  1. Holmgren, A. (1985) Annu. Rev. Biochem. 54:237.
  2. Powis, G. and W.R. Monfort (2001) Annu. Rev. Pharm. Toxicol. 41:269.
  3. Deiss, L.P. and A. Kimchi (1991) Science 252:117.
  4. Spyrou, G. et al. (1997) J. Biol. Chem. 272:2936.
  5. Miranda-Vizuete, A. et al. (1998) Biochem. Biophys. Res. Commun. 243:284.
  6. Nakamura, H. et al. (1997) Annu. Rev. Immunol. 15:147.
  7. Mourice, M.M. et al. (1999) Arthritis Rheum. 42:2430.
  8. Jikimoto, T. et al. (2001) Mol. Immunol. 38:765.
Entrez Gene IDs
7295 (Human)
Alternate Names
Thioredoxin1; Thioredoxin-1; Trx1; TXN; TXN1

Citations for Recombinant Human Thioredoxin-1 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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  1. Gram Negative Bacterial Inflammation Ameliorated by the Plasma Protein Beta 2-Glycoprotein I
    Sci Rep, 2016;6(0):33656.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  2. Methodological aspects of ELISA analysis of thioredoxin 1 in human plasma and cerebrospinal fluid.
    Authors: Lundberg M, Curbo S, Reiser K, Masterman T, Braesch-Andersen S, Arestrom I, Ahlborg N
    PLoS ONE, 2014;9(7):e103554.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA (Standard)
  3. A human serum albumin-thioredoxin fusion protein prevents experimental contrast-induced nephropathy.
    Authors: Kodama A, Watanabe H, Tanaka R, Tanaka H, Chuang V, Miyamoto Y, Wu Q, Endo M, Hamasaki K, Ishima Y, Fukagawa M, Otagiri M, Maruyama T
    Kidney Int, 2013;83(3):446-54.
    Species: Rat
    Sample Types: In Vivo
    Applications: Bioassay
  4. Redox control of beta2-glycoprotein I-von Willebrand factor interaction by thioredoxin-1.
    Authors: Passam FH, Rahgozar S, Qi M, Raftery MJ, Wong JW, Tanaka K, Ioannou Y, Zhang JY, Gemmell R, Qi JC, Giannakopoulos B, Hughes WE, Hogg PJ, Krilis SA
    J. Thromb. Haemost., 2010;8(8):1754-62.
    Species: Human
    Sample Types: Buffer
    Applications: Enzyme Assay

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