This antibody reacts with E. coli Trx and Trx-tag-fusion protein. I detects 12KD of E.coli TrxA by Western blotting on total cell lysate from induced E. coli harboring Trx-expression vector.
Western blot: 1:1000 for chemiluminescence detection system. Can be used for West-Western blot.
实验流程
1) Boil all samples for 3~5 minutes. Load 10 μL of cell lysate or tissue homogenate (5~20 μg total protein) to each well of an SDS - polyacrylamide gel and electrophorese in a 1 mm thick gel. 2) Transfer to a p olyvinylidene difluoride (PVDF) membrane at 10V f or 1 hour in a semi-dry transfer system. (Transfer Bu ffer: 25 mM Tris, 190 mM glycine, 20 % MeOH). 3) The transferred protein s can be visualized by staining the membrane for 1 minute with Ponceau S. Rinse the membrane with PBS. 4) Non-specific binding sites are blocked by immersing the membrane in Block Ace TM for 1 hour at room t emperature, or for overnight at 4 ° C. 5) Incubate in primary an t ibody diluted as suggested in the APPLICATIONS for 1 hour at room temperature. (The optimal antibody concentration will depend on the experimental variables and the abundance of the antigen.) 6) Wash the membrane 3 times with PBS, 0.2 % Tween20 for 5~10 minutes per wash. 7) Incubate in Horseradish Peroxidase conjugated goat anti-mouse diluted 1:10 000 in PBS, 0.2 % Tween20 for 45 minutes at room temperature. 8) Wash the membrane 3 times with PBS, 0. 2 % Tween20 for 10 minutes per wash. 9) Incubate in Ame rsham ECL Reagent for 1 minute. Drain membrane, remove excess ECL Reagent by dabbing with a Kimwipe, and seal in plastic wrap. 10) Expose to ECL hyperf ilm in a dark room for 30 seconds. Develop as usual for autoradiogram or X-ray. The conditions for development and exposure may vary.
限制
仅限研究用
状态
Liquid
缓冲液
PBS containing 50 % glycerol. Contains no preservatives
储存液
Without preservative
储存条件
-20 °C
储存方法
Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: One year from despatch.
抗原
TXN
(Thioredoxin (TXN))
别名
thioredoxin,trx1
背景
The thioredoxin (Trx) fusion E. coli expression system is avail a ble to offer soluble expression of normally insoluble o r difficult to express proteins. LaVallie et al. 1) reported that a number of mammalian cytokines and growth factors, when expressed as C-terminal trxA fusion proteins, stayed remarkably soluble in the E. coli cytoplasm under certain conditions.