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Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Thioredoxin/ TRX.
(Peptide available as ab27815.)
Our Abpromise guarantee covers the use of ab26320 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 12 kDa (predicted molecular weight: 12 kDa).|
|ICC/IF||1/200 - 1/1000.|
ab26320 staining TRX in HLE-Be lens cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 3.7% formaldehyde, permeabilized with 0.25% Triton X-100 in PBS and blocked with 1% BSA. Samples were incubated with primary antibody (1/1500) overnight at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody (1/1000). TRX stained in green, Mitotracker red and merging of the 2 images.
ab26320 staining Thioredoxin (red) in Cat urinary bladder tissue by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were blocked with 1% BSA in TBS-T for 40 minutes at room temperature and heat antigen retrieval was processed at 99°C for 15 minutes in 0.1M EDTA-NaOH buffer. Samples were incubated with primary antibody (1/100) overnight at 4°C. Nuclei stained with DAPI (blue). a = Transitional cell epithelium, b = Lamina propria mucosae, c = Muscle tunic
Thioredoxin / TRX was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Thioredoxin / TRX and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab26320.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 12kDa; Thioredoxin / TRX
ab26320 detects a band of 12 kDa in HeLa, Jurkat and A431 whole cell lysates. The data for the Jurkat and A431 lysates is not shown.
Immunofluorescent detection of Thioredoxin /TRX using antibody ab26320. Chinese Hamster Cell (Ovary cells) were fixed in formaldehyde and permeabilized in 0.1% Triton X in PBS/3% BSA. Primary antibody was incubated at 1/1000 for 1h @ 4°C in PBS containing 3% BSA. Secondary antibody: anti-rabbit conjugated to Alexa Fluor® 568 used @ 1/300. CHO cells co-transfected with plasmids encoding GFP and a mutated form of human thioredoxin which does not enter the nucleus, and labelled using ab26320 (1/1000) and Alexa 568 (1/300). Panel [A] demonstrates GFP immunoreactivity. [B ]thioredoxin immunoreactity; many of the same cells are immunoreactive for both GFP and thioredoxin. Panel [D] thioredoxin primary antibody was omitted, hence there is no immunoeactivity.
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