Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab43014 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
IHC-Fr Use at an assay dependent concentration. PubMed: 20651230
ChIP Use at an assay dependent concentration. PubMed: 22073215
CHIPseq Use at an assay dependent concentration. PubMed: 22073215
WB Use at an assay dependent concentration. Predicted molecular weight: 14 kDa.
ELISA Use at an assay dependent concentration.

Target

  • Relevance
    HIV1 tat (Transactivator of Transcription) protein is a pleiotropic factor that induces a broad range of biological effects in numerous cell types. At the HIV promoter, tat is a powerful transactivator of gene expression which acts by inducing chromatin remodeling and by recruiting elongation-competent transcriptional complexes on to the viral LTR.
  • Cellular localization
    Nuclear. Upon stimulation with EDN1, it is exported from the nucleus to the perinuclear region.
  • Alternative names
    • p14 antibody
    • Protein Tat antibody
    • Transactivating regulatory protein antibody

Images

  • All lanes : Anti-HIV1 tat antibody - ChIP Grade (ab43014) at 1/2000 dilution

    Lane 1 : Jurkat whole cell lysate 0 hours after infection
    Lane 2 : Jurkat whole cell lysate 2 hours after infection
    Lane 3 : Jurkat whole cell lysate 4 hours after infection
    Lane 4 : Jurkat whole cell lysate 6 hours after infection
    Lane 5 : Jurkat whole cell lysate 8 hours after infection
    Lane 6 : Jurkat whole cell lysate 10 hours after infection

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : HRP Goat anti-rabbit IgG polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 14 kDa
    Observed band size: 14 kDa



    Jurkat cells were pNL4.3 infected and samples taken at 2 hour intervals for blotting.

    See Abreview

  • Immunohistochemical analysis of normal mouse brain, labeling HIV1 tat with ab43014. Samples were fixed in 4% paraformaldehyde overnight, cryoprotected in 30% sucrose and frozen at -80°C. 10µm thick frozen coronal sections were blocked in 5% fetal bovine serum, 1% Triton X-100, 0.5% Tween 20 and 1% skim milk in PBS for 2 hours at RT. Samples were incubated with ab43014 overnight at 4°C, and an alexa fluor conjugated secondary antibody for 2 hours at RT. 

References

This product has been referenced in:
  • Lee FH  et al. Disrupting GluA2-GAPDH Interaction Affects Axon and Dendrite Development. Sci Rep 6:30458 (2016). IHC-P ; Mouse . Read more (PubMed: 27461448) »
  • Romani B  et al. HIV-1 Vpr reactivates latent HIV-1 provirus by inducing depletion of class I HDACs on chromatin. Sci Rep 6:31924 (2016). Read more (PubMed: 27550312) »
See all 12 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T cells)
Gel Running Conditions
Reduced Denaturing (4-12)
Loading amount
10 µg
Treatment
Transfected with HIV-1 NL4-3 for 48 hours
Specification
HEK293T cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 19 2016

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (gel 15%)
Sample
Human Cell lysate - whole cell (astrocyte U251)
Specification
astrocyte U251
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 Oct 15 2013

Answer

Thank you very much for your email. My colleague is out of the office but I'll be happy to assist you with this issue.

I've looked at our stock, and unforunately we only have the same lot that did not work well for you. Did the replacement vial that my colleague sent work as expected?

I apologize for the inconvenience, but at this time we are not sure when a new lot will be produced.

I hope this will be useful, but please let me know if you have further questions and I'll be happy to help.

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of the old lotwith the order number. This is shipping from our UK office, so you can expect it around Sept 11th. To check the status of the order please contact our Customer Service team and reference this number.

Your credit note ID for the other vial is XXXXX. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Read More

Answer

Thank you for your reply and for providing the additional information.

It appears that the antibody isn't working as expected, so I would like to offer you a free of charge replacement or credit note. We only have the same lot in stock in our US office as the one you tried that didn't work well. We do have 1 vial of the old lot that worked well GR65428-5, but this would ship from our UK office and will take a few extra days.

How would you like to proceed? Would you like us to send you a replacement vial from the old lot that worked well?

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Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

What is the problem that you are experiencing? Is it no bands in WB or enrichment in ChIP? I am listing some questions below so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

1) Abcam product code ab43014

2) Abcam order reference number or product batch number GR87684-1

3) Description of the problem

4) Sample preparation:

Type of sample (whole cell lysates, fraction, recombinant protein…):

Lysis buffer

Protease inhibitors:

Phosphatase inhibitors

Reducing agent

Boiling for ≥5 min? yes/no

Protein loaded ug/lane or cells/lane

Positive control

Negative control

5) Percentage of gel

Type of membrane

Protein transfer verified

Blocking agent and concentration

Blocking time

Blocking temperature

6) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution

Diluent buffer

Incubation time

Incubation temperature:

7) Secondary antibody:

Species:

Reacts against:

Concentration or dilution

Diluent buffer

Incubation time

Incubation temperature:

Fluorochrome or enzyme conjugate:

8) Washing after primary and secondary antibodies:

Buffer

Number of washes

9)Detection method

10) How many times have you run this staining?

Do you obtain the same results every time?

What steps have you altered to try and optimize the use of this antibody?

Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

I look forward to receiving your reply.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Jurkat Cells)
Loading amount
25 µg
Specification
Jurkat Cells
Treatment
pNL4.3 infected
Gel Running Conditions
Reduced Denaturing (15% SDS-PAGE gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 01 2012

Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.



I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1048500.


To check the status of the order please contact our Customer Service team and reference this number.


Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.



I wish you the best of luck with your research.

Read More

Question
Answer

DISCOUNT CODE: **********
Expiration date: **************

I am very pleased to hear you would like to accept our offer and test ab43014 in ICC/IF. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview forICC/IF and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for your patience. I have enquired at the originator of the product but unfortunately, this product has not been tested against human macrophages and HIV strain BAL. Therefore, we do not know if it will be suitable for your research. If you decide to go ahead and purchase this product, we encourage you to submit an Abreview via the online product datasheet. We always encourage customers to send their results back to us, whether positive or negative, and we make all product information available to researchers. To find out more about our Abreview system, please see the following URL: https://www.abcam.com/abreviews I am sorry I can't be of more assistance in this occasion but please do not hesitate to contact us if you need any more advice or information.

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

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