Overview

  • Product name

    Anti-Thioredoxin / TRX antibody
    See all Thioredoxin / TRX primary antibodies
  • Description

    Rabbit polyclonal to Thioredoxin / TRX
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Sheep, Horse, Cow
  • Immunogen

    Recombinant human protein purified from E.coli.

  • Positive control

    • IHC-P: FFPE human ovary tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab86255 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/2000. Predicted molecular weight: 12 kDa.
ICC/IF 1/200.
IHC-P 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function

    Participates in various redox reactions through the reversible oxidation of its active center dithiol to a disulfide and catalyzes dithiol-disulfide exchange reactions. Plays a role in the reversible S-nitrosylation of cysteine residues in target proteins, and thereby contributes to the response to intracellular nitric oxide. Nitrosylates the active site Cys of CASP3 in response to nitric oxide (NO), and thereby inhibits caspase-3 activity. Induces the FOS/JUN AP-1 DNA-binding activity in ionizing radiation (IR) cells through its oxidation/reduction status and stimulates AP-1 transcriptional activity.
    ADF augments the expression of the interleukin-2 receptor TAC (IL2R/P55).
  • Sequence similarities

    Belongs to the thioredoxin family.
    Contains 1 thioredoxin domain.
  • Post-translational
    modifications

    In the fully reduced protein, both Cys-69 and Cys-73 are nitrosylated in response to nitric oxide (NO). When two disulfide bonds are present in the protein, only Cys-73 is nitrosylated. Cys-73 can serve as donor for nitrosylation of target proteins.
    In case of infection, ubiquitinated by S.typhimurium protein slrP, leading to its degradation.
  • Cellular localization

    Nucleus. Cytoplasm. Secreted. Secreted by a leaderless secretory pathway. Predominantly in the cytoplasm in non irradiated cells. Radiation induces translocation of TRX from the cytoplasm to the nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • ADF antibody
    • ATL derived factor antibody
    • ATL-derived factor antibody
    • DKFZp686B1993 antibody
    • MGC61975 antibody
    • SASP antibody
    • Surface associated sulphydryl protein antibody
    • Surface-associated sulphydryl protein antibody
    • testicular tissue protein Li 199 antibody
    • THIO_HUMAN antibody
    • Thioredoxin antibody
    • thioredoxin delta 3 antibody
    • TRDX antibody
    • TRX 1 antibody
    • Trx antibody
    • TRX1 antibody
    • TXN antibody
    • TXN delta 3 antibody
    • TXN protein antibody
    • zgc:92903 antibody
    see all

Images

  • ab86255 staining Thioredoxin by Western blot.
    Lane 1: HeLa cell lysate
    Lane 2: A431 cell lysate
  • IHC image of Thioredoxin / TRX staining in human ovary formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab86255, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ICC/IF image of ab86255 stained HeLa cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab86255, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:

  • Danzl K  et al. Early inhibition of endothelial retinoid uptake upon myocardial infarction restores cardiac function and prevents cell, tissue, and animal death. J Mol Cell Cardiol 126:105-117 (2019). Read more (PubMed: 30472251) »
  • Zhu J  et al. Oxidative and anti-oxidative status in muscle of young rats in response to six protein diets. Sci Rep 7:13184 (2017). Read more (PubMed: 29030561) »
See all 11 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 29 2016

Application
Western blot
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 26 2016

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (15% Gel)
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 05 2014

Answer

Thank you foryour reply.

I have not been able to link any of our antibodies to the one used in this article, did they suggest a catalogue number?

We have several antibodies against thioredoxin which may be of interest to you. The only one we have which we know works in staining of frozen sections is the mouse monoclonal antibody https://www.abcam.com/Thioredoxin-TRX-antibody-2B1-ab1754.htmlwhich you have made a reference to. In answer to your question, this antibody has been shown to also detect the mouse thioredoxin protein.

We have several other antibodies which although they have not been tested with frozen sections as yet, may be suitable for this application. However as we have not tested it ourselves we would not be able to guarantee this:

https://www.abcam.com/Thioredoxin-TRX-antibody-ab26320.html

https://www.abcam.com/Thioredoxin-TRX-antibody-ab86255.html

These are both rabbit polyclonals which have been shown to detect both the mouse and human thioredoxin.

If you would like to try any of these antibodies in IF staining of mouse or human tissue I would be able to offer you a testing discount.

This offer means that if you purchaseany of the antibodies mentioned aboveas normal, test the antibody with yourmouse or human tissue in IF and let us know of the results through an Abreview (no matter whether positive or negative) you would be eligible for a free primary antibody of your choice from our catalogue (or the equivalent price of the antibody purchased off any product). More information on this offer can be found here:

www.abcam.com/collaborationdiscount

Please note that the antibody to be tested must be purchased, tested, the Abreview submitted and the free product claimed within a 4 month period.

If you would be interested in participating in this scheme please do let me know as a discount code needs to be issued prior to the purchase of the antibody to be tested.

I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

Read More

Answer

Thank you for your reply.

Under our Abpromise we do guarantee our products to work as stated on the datasheet for 6 months and if you experience any problems with it during that time frame then and we cannot resolve the issue you are having then we would be happy to either send a replacement antibody or to process a refund.

I understand that you are experienced in western blotting techniques but there is always chance that we could spot something that will help resolve the issue, so I would be very grateful if you could answer the questions below:.

1 - How much sample are you loading?

2 - What primary antibody concentrations have you tried?

3- How long do you boil your samples for? As possible the protein can be complexed with other proteins and so boiling longer may break up this complex.

4 - In your original email, you mentioned you got "very dark non-specific bands", did you get more than one band on the membrane?

5 - Have you tried blocking in 5% milk rather than BSA?

6 - How was the antibody stored?

I look forward to your reply and helping you get this antibody working correctly in western blot.

Read More

Answer

Thank you for contacting Abcam.

I am sorry that you are experiencing problems with ab16835 in western blot and I would be more than happy to help you troubleshoot these problems.

We removed ab16835 from our catalogue in 2009 and so I was wondering when you purchased the antibody and also if you experienced any issues with it when it first arrived?

Also, if you could provide me with a copy of the protocol that you are using, I will be very happy to look it over and see if there are any suggestions that I can make (unfortunately your attachment did not come through, so it would be greatly appreciated if you could resend this).

I look forward to your reply and helping you improve your results.

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (lens epithelial cells)
Loading amount
25 µg
Specification
lens epithelial cells
Gel Running Conditions
Reduced Denaturing (12%SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Dec 22 2011

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up