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: IP, WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Cat, Human
    Predicted to work with: Rat, Sheep, Cow, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Thioredoxin/ TRX.

    Read Abcam's proprietary immunogen policy (Peptide available as ab27815.)

  • Positive control
    • Recombinant human Thioredoxin / TRX protein (ab51064) can be used as a positive control in WB. HeLa, Jurkat and A431 whole cell lysates.

Properties

Applications

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.

Application Abreviews Notes
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.
IHC-P 1/2000.

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

  • Anti-Thioredoxin / TRX antibody (ab26320) at 1 µg/ml + Recombinant human Thioredoxin / TRX protein (ab51064) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 12 kDa


    Exposure time: 30 seconds
  • 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

  • Anti-Thioredoxin / TRX antibody (ab26320) at 1 µg/ml + HeLa whole cell lysate at 20 µg

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

    Performed under reducing conditions.

    Predicted band size: 12 kDa
    Observed band size: 12 kDa



    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.

     

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

    See Abreview

  • 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.

    See Abreview

  • ab26320 staining Thioredoxin/TRX in Human lung tissue sections by IHC-P (Formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5 minutes of peroxidase block followed by 10 minutes of protein block at 20°C; antigen retrieval was heat mediated in retrieval solution. Samples were incubated with primary antibody (1/2000 in antibody diluent) for 45 minutes at 20°C. An undiluted HRP-conjugated polymer goat anti-mouse/rabbit IgG polyclonal polymer was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Han Y  et al. Reactive oxygen species promote tubular injury in diabetic nephropathy: The role of the mitochondrial ros-txnip-nlrp3 biological axis. Redox Biol 16:32-46 (2018). Read more (PubMed: 29475133) »
  • Stancic A  et al. A lesson from the oxidative metabolism of hibernator heart: Possible strategy for cardioprotection. Comp Biochem Physiol B Biochem Mol Biol 219-220:1-9 (2018). WB . Read more (PubMed: 29501789) »
See all 12 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (Hep G2)
Specification
Hep G2
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Dec 26 2014

Answer

Without reduced and denatured samples, since the antibody was raised against a peptide it may not be able to recognize its epitope under the non-reduced, non-denatured conditions of the redox WB.

Since you're samples in the redox WB are non-reduced and non-denatured, if you know the structure of the protein then you might then be able to determine if the immunogen sequence is likely to be present in the interior or on the surface of the protein and how likely it is to be accessible.

Read More

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

Merci de votre intérêt pour nos anticorps. La TRX80 correspond effectivement aux 80 acides aminés N terminaux de TRX. J'ai donc cherché dans nos anticorps si nous avons un qui reconnait un épitope dans les premiers 80 acides aminés. Pour la plupart des anticorps TRX, je ne peux pas conclure, car l'anticorps était produit contre la protéine entière. Pour le ab26320 et ab109385, nous savons qu'il sont que contre la forme longue, car l'immunogène est C terminal, après le acide aminé 80. Le ab26928 lui par contre, reconnait la forme longue et forme courte, car l'immunogène correspond aux acides aminés 1-60 de la TRX humaine. Le ab26320 et ab109385 sont testés en WB et contre la protéine de souris. Si vous tester un de ces anticorps aussi en ELISA, je serais ravi de vous faire la même offre de test que pour e ab86983 ci-dessous. Le ab26928 est testé en WB, mais pas encore contre la protéine de souris. Je peux aussi vous faire la même offre de test que pour le ab86983 pour tester le ab26928 sur la protéine de souris. L'alignement entre la protéine TRX humaine et de souris laisse penser qu'il y a des chances raisonnables de crossréactivité de cet anticorps. http://www.uniprot.org/align/201109192CDUIL3Q6A Comme nous n'avons pas encore testé le ab86983 sur du tissue de souris, il me sera possible de vous offrir une remise si vous souhaitez tester ceci vous même. Après soumission d’une Abreview avec vos résultats, une remise correspondant à la valeur d’achat de ab86983 pourra être utilisable lors d’une prochaine commande d’anticorps primaire. Etapes à suivre : 1. Nous confirmer que vous souhaiteriez tester ab86983 chez la souris afin de recevoir le code de remise correspondant. Ce code doit être édité avant l’achat de ab86983 2. Acheter ab86983 par téléphone, fax ou internet (www.abcam.com) 3. Le tester chez la souris 4. Nous faire part de vos résultats, positifs ou négatifs, grâce à notre système Abreview. Pour plus d’informations https://www.abcam.com/abreviews 5. Après soumission de votre Abreview, appeler notre service clientèle afin de passer votre prochaine commande grâce au code de réduction. Ce code de réduction est utilisable 120 jours après son édition et utilisable lors de l’achat d’un autre anticorps primaire. Même si ab86983 est démontré ne pas fonctionner chez la souris, après soumission de votre Abreview, vous recevrez quand même une remise sur l’achat de votre prochain anticorps. N’hésitez pas à nous demander plus d’informations concernant cette offre promotionnelle. Termes et Conditions : www.abcam.com/collaborationdiscount.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Lung)
Specification
Lung
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Target Retreival Solution
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C

Mr. Antibody Solutions

Verified customer

Submitted Mar 22 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Chinese Hamster Cell (Ovary cells)
Specification
Ovary cells
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% Triton X in PBS/3% BSA

Mr. Carl Hobbs

Verified customer

Submitted Feb 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Dec 18 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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