Anti-Thioredoxin / TRX antibody

• ICC/IF

AbReview6300****

Immunocytochemistry/ Immunofluorescence - Anti-Thioredoxin / TRX antibody (AB26320)

ab26320, at 1/200 dilution, staining human TRX in assynchronous HeLa cells by ICC/IF.

This image is courtesy of an Abreview submitted by Dr Kirk McManus

• IP

Unknown

Immunoprecipitation - Anti-Thioredoxin / TRX antibody (AB26320)

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

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Immunoprecipitation - Anti-Thioredoxin / TRX antibody (ab26320)

Predicted band size: 12 kDa

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Exposure time: 20min

• WB

Project1649****

Western blot - Anti-Thioredoxin / TRX antibody (AB26320)

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.

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Western blot - Anti-Thioredoxin / TRX antibody (ab26320) at 1 µg/mL

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HeLa whole cell lysate at 20 µg

Secondary

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Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

Predicted band size: 12 kDa

Observed band size: 12 kDa

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• ICC/IF

AbReview17259****

Immunocytochemistry/ Immunofluorescence - Anti-Thioredoxin / TRX antibody (AB26320)

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.

This image is courtesy of an abreview submitted by Dr David Tonge, Wolfson CARD, Kings College London

• WB

Unknown

Western blot - Anti-Thioredoxin / TRX antibody (AB26320)

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Western blot - Anti-Thioredoxin / TRX antibody (ab26320) at 1 µg/mL

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Western blot - Recombinant human Thioredoxin / TRX protein (Active) (<a href='/en-us/products/proteins-peptides/recombinant-human-thioredoxin-trx-protein-active-ab51064'>ab51064</a>) at 0.01 µg

Secondary

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Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 12 kDa

true

Exposure time: 30s

• WB

CiteAb

Western blot - Anti-Thioredoxin / TRX antibody (AB26320)

Thioredoxin / TRX western blot using anti-Thioredoxin / TRX antibody ab26320. Publication image and figure legend from Dehghan, E., Zhang, Y., et al., 2017, Nat Commun, PubMed 29263362.

ab26320 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab26320 please see the product overview.

Hydralazine enhances NRF2 signaling in SH-SY5Y cells. a Hydralazine increased cellular NRF2 protein in a dose-dependent manner demonstrated by western blot analysis of cell lysates (input) and NRF2 immunoprecipitates (IP : NRF2). *p < 0.05 and **p < 0.01, two-tailed Student's t test, n = 3, mean ± SD. b The mass spectrometry based label-free quantification of NRF2 immunoprecipitates prepared in part A. Extracted ion chromatogram of a NRF2 peptide, IINLPVVDFNEMoxMoxSK, as well as bar plot quantification are shown. Samples were treated with 0.05% H2O2 prior to mass spectrometric analysis. *p < 0.05, two-tailed Student's t test, n = 3, mean ± SD. c Hydralazine reduced the interaction between NRF2 and KEAP1. Interactions were measured by reciprocal Co-IPs followed by western blot analysis. *p < 0.05, two-tailed Student's t test, n = 3, mean ± SD. d NRF2 translocates to the nucleus with hydralazine treatment. Treated cells were subjected to cell fractionation and western blot analysis. *p < 0.05 and **p < 0.01, two-tailed Student's t test, n = 3, mean ± SD. e Hydralazine treatment increased nuclear NRF2 phosphorylation quantified using an antibody specific to NRF2 phosphorylation at serine 40. *p < 0.05 and **p < 0.01, two-tailed Student's t test, n = 3, mean ± SD. f TXN, a potent regulator of the NRF2–KEAP1 response system, is upregulated with hydralazine treatment. *p < 0.05, two-tailed Student's t test, n = 3, mean ± SD. g ARE-driven luciferase activity was increased with hydralazine treatment, indicating an increase in the transcriptional activation of NRF2 target genes. Hydrazine did not increase luciferase activity. **p < 0.01, two-tailed Student's t test, n = 6, mean ± SD. h Hydralazine treatment increased the expression of NRF2 downstream targets measured by qPCR using actin as internal control. For the list of primers used for qPCR, see Supplementary Table 1 online. *p < 0.05, and **p < 0.01, two-tailed Student's t test, n = 6, mean ± SEM. i Hydralazine treatment increased the expression of NRF2 downstream target measured by western blot analysis. *p < 0.05, two-tailed Student's t test, n = 3, mean ± SD

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