TXN antibody

Catalog No. ABIN5540698

This anti-TXN antibody is a Mouse Monoclonal antibody detecting TXN in WB. Suitable for Human.

Quick Overview for TXN antibody (ABIN5540698)

Target: TXN (Thioredoxin (TXN))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This TXN antibody is un-conjugated

Application: Western Blotting (WB)

Clone: 2E3

Product Details anti-TXN Antibody

Specificity: This antibody reacts with thioredoxin.

Cross-Reactivity (Details): Does not treact with Mouse (NIH/3T3, WR19L, Ba/F3) and Rat (PC12).

Purification: Protein A agarose

Immunogen: Recombinant full-length human thioredoxin

Isotype: IgG2a

Application Details

Application Notes: Western blot: 1 mg/mL for chemiluminescence detection system. For details see protocol below.

Protocol: SDS-PAGE & Western Blotting 1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mM Tris-HCl, pH 7.2, 250 mM NaCl, 0.1 % NP-40, 2 mM EDTA, 10 % glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. Measure the protein concentration of the supernatant and add the cold Lysis buffer to make 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli's sample buffer. 4) Boil the samples for 2 minutes and centrifuge. Load 10 μ L of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in a semi-dry transfer system (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20 % MeOH). See the manufacture's manual for precise transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10 % skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature, or overnight at 4 o C. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7.2 containing 1 % skimmed milk as suggest in the APPLICATIONS for 1 hour at room temperature. (The concentration of antibody will depend on condition.) 8) Wash the membrane with PBS-T [0.05 % Tween-20 in PBS] (5 minutes x 6 times). 9) Incubate the membrane with the 1:10,000 HRP-conjugated anti-mouse IgG diluted with 1 % skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 10) Wash the membrane with PBS-T (5 minutes x 6 times). 11) Wipe excess buffer on the membrane, then incubate it with appropriate chemilumin escence reagent for 1 minute. Remove extra reagent from the membrane by dabbing with paper towel, and seal it in plastic wrap. 12) Expose to an X-ray film in a dark room for 5 minutes. Develop the film as usual. The condition for exposure and development may vary. (Positive controls for Western blotting Raji, HeLa)

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: PBS containing 50 % glycerol, pH 7.2. No preservative is contained.

Preservative: Azide free

Storage: -20 °C

Storage Comment: Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: One year from despatch.

Target Details for TXN

Target: TXN (Thioredoxin (TXN))

Alternative Name: thioredoxin,trx1

Background: Thioredoxin (TRX) is a small ubiquitous protein (12 kDa) which is exist in a wide activity of prokaryotic and eukaryotic cells. TRX is activated by reducing its disulfide group (-S-S- → 2SH) between cysteine residues within the conserved active site sequence: -Cys-Gly-Pro-Cys-. The reduction of TRX is specifically catalyzed by "TRX reductase", a FAD and selenocystein containing protein, using NADPH. Once reduced, TRX in turn reduces disulfide bonds of various proteins to regulate the activity of the protein (redox regulation). TRX has multiple biological function Activation of transcripti on factors such as NF- κ B and glucocotricoid receptor, anti-apoptotic function, stimulation of cytokine expression, tissue protection from ischemia-reperfusion injury, cytoprotection from cytotoxic and DNA-damaging agents, proliferation, and drug resistance in certain malignancies. A part of these functions are due to TRX which exported out of the cell where it has additional ability to stimulate cell growth, cytoprotection, and co-cytokine activity. It is said that some of above TRX functions are related to response against the cellular "stresses". In fact, TRX is translocated from the cytosol into the nucleus and/or highly expressed by various stress such as UV irradiation and viral infection. So, it is suggested that TRX may have a role in the cellular responses against the stresse s through the regulation of intracellular redox (oxidation-reduction) atatus.

UniProt: P10599

Pathways: Carbohydrate Homeostasis, Cell RedoxHomeostasis