Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab6539 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
Dot blot 1/3000.
WB 1/3000. Predicted molecular weight: 37 kDa.
IP 1/750.

Target

  • Relevance
    The transcriptional transactivator (Tat) is a key regulatory protein of HIV. It is expressed early after the virus integrates into the cell, and stimulates the elongation of RNA polymerase II. It binds onto a sequence known as the TAR, or transactivator response element, located at the end of the HIV genetic chain. There, the tat protein helps assemble new copies of HIV. The tat protein-TAR complex speeds up the rate of viral reproduction by about a thousand times. If it is not present, the transcription process frequently stops short, and few functional HIV particles are produced. Tat is an important potential target for antiretrovirals and vaccine development.
  • Alternative names
    • p14 antibody
    • Tat antibody
    • Tat protein antibody
    • Transactivating regulatory protein antibody

References

This product has been referenced in:
  • Midde NM  et al. Mutation of tyrosine 470 of human dopamine transporter is critical for HIV-1 Tat-induced inhibition of dopamine transport and transporter conformational transitions. J Neuroimmune Pharmacol 8:975-87 (2013). Read more (PubMed: 23645138) »
See 1 Publication for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Neurons)
Permeabilization
Yes - PBS Triton X-100
Specification
Neurons
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 12 2018

Answer

Thank you for contacting us. Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase. However, in this case we wouldn't be able to guarantee the antibody will work to bind your peptide bound to an enzyme.

I would probably recommend the polyclonal rather than the monoclonal since the polyclonal will have multiple binding epitopes. However, ab6539 hasn't been tested in immunohistochemistry. If you would like to test this antibody in IHC, let me know as you would qualify for our testing discount program. Essentially you would purchase this antibody, test it in IHC, submit an Abreview with your data (whether positive or negative) and receive a discount code for a free antibody off your next Abcam purchase within 4 months.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

Read More

Answer

Thank you for contacting us.

The immunogen for ab24778 is within the first 37 amino acids of human HIV TAT protein. For ab6539, the immunogen is full length TAT protein tagged with a GST fusion protein. However, we have not mapped the exact epitope for this antibody.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question

BATCH NUMBER 64866 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Non-specific band This antibody recognizes HIV rev. In 293T cells transfected with a HIV rev only expressing plasmid, this antibody clearly bound to rev in Western blot. In cells transfected with plasmids expressing HIV rev and tat, this antibody only bound to HIV rev. I was unable to visualize any banding pattern for HIV tat. I have repeated this 3 times. SAMPLE human/whole cell extract/expressed from plasmid PRIMARY ANTIBODY ab6539 1:3000 1hr room temp./TBS-0.5% Tween 20 5%BSA/1hr. room temp/ 3 washes 5min each with plenty of TBS-1%Tween 20. SECONDARY ANTIBODY JacksonImmunoReserach labs/donkey anti-sheep HRP/1:40,000/1hr/room temp/ 3 washes 5min each with plenty of TBS-1%Tween 20. DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED 293T cells tranfected with plasmids known to express HIV tat and rev, HIV rev, cells infected with virus expressing HIV tat and rev. Virus preps verified to express HIV tat as determined by lacZ staining, only possible if HIV tat is expressed from virus. cells infected with about 4x108 iu/ml ANTIBODY STORAGE CONDITIONS Aliquot antibody in 12microliters/tube stored at -20C SAMPLE PREPARATION 50mM Tris pH8.0, 150mM NaCl, 1mM EDTA, 1% TritonX-100/Roche complete mini tablet. (1 tablet/10mls of buffer), freeze thaw several times, pellet debris, mix sample with Bio-rad XT 5x sample buffer and 20x reducing agent, heat 95C 5min. AMOUNT OF PROTEIN LOADED 100micrograms/lane ELECTROPHORESIS/GEL CONDITIONS Bio-rad precast gel, XT Critterion 12% gel, 200volts, 45-60mins. using MOPS buffer from Bio-rad. TRANSFER AND BLOCKING CONDITIONS Wet transfer, tris-glycine buffer, 30min. 100volts, PVDF, block using 5%BSA 1hr. room temp HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES There is no reason for HIV tat antibody to recognize HIV rev.

Read More
Answer

I am very sorry for the delay in responding to you. The source of the antibody told us: "This concern is a little puzzling, because this antisera is from a large pool that we have been selling consistently since the 1990’s. This antisera has been stored frozen the entire time. As you know, the specificity of an antibody does not change over time. According to our records, we have never received any complaints claiming that this antibody is reactive with rev and not tat. Quite the contrary, we have many customers that have been repeatedly purchasing this product for years. As the data sheet states, antibody specificity was analyzed by immunoblotting with control TAT fusion protein (MW 37 kD) with detection using the ECL system. The antibody will detect wild type HIV-TAT protein. The portion of TAT used for the immunogen was amino acids 49-57. This antibody was also tested by immunoprecipitation using a dilution of 1:750 and protein G as the immnoprecipitant. 20-HS75 recognized P-32 labelled HIV-TAT very well in this assay." In this instance, without admission of responsibility or liability, I am prepared to offer you a refund or credit note. Please let me know which you would prefer and the Abcam order number and/or PO number and I will arrange accordingly.

Read More

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

Sign up