Overview

  • Product name

    Anti-Torsin A/DYT1 antibody
    See all Torsin A/DYT1 primary antibodies
  • Description

    Rabbit polyclonal to Torsin A/DYT1
  • Host species

    Rabbit
  • Specificity

    The immunogen used to raise this antibody has 92% homology with the human Torsin A/DYT1 protein. Some customers have successfully used ab34540 in human tissue. Please contact Abcam Scientific Support for more information.

  • Tested applications

    Suitable for: IHC-FoFr, ICC/IF, WBmore details
  • Species reactivity

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

    Synthetic peptide corresponding to Rat Torsin A/DYT1 aa 300 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab35510, ab35511)

  • Positive control

    • This antibody gave a positive control in the following mouse lysates: Brain tissue Testis tissue Heart tissue Skeletal Muscle tissue Liver tissue Mouse Fetal lysate - 14 Days old Tissue Brain Tissue Lysate - normal tissue, 0 days old NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell MEF1 (Mouse embryonic fibroblast cell line) Whole Cell This antibody gave a positive control in the following rat lysates: Liver tissue PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell
  • General notes

     This product was previously labelled as Torsin A

     

Properties

Applications

Our Abpromise guarantee covers the use of ab34540 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-FoFr 1/100.
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 38 kDa (predicted molecular weight: 38 kDa).

Target

Images

  • All lanes : Anti-Torsin A/DYT1 antibody (ab34540) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Mouse Brain at 10 µg
    Lane 3 : Testis (Mouse) Whole Cell Lysate - normal tissue at 10 µg
    Lane 4 : Mouse Heart Mouse Tissue Lysate at 10 µg
    Lane 5 : Mouse skeletal muscle tissue lysate - total protein (ab29711) at 10 µg
    Lane 6 : Mouse Fetal lysate - 14 Days old Tissue Lysate at 10 µg
    Lane 7 : Mouse brain tissue lysate - total protein (0 days) (ab7188) at 10 µg
    Lane 8 : Mouse Embryonic Fibroblast 1 at 10 µg
    Lane 9 : NIH 3T3 whole cell lysate (ab7179) at 10 µg

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution

    Performed under reducing conditions.

    Predicted band size: 38 kDa
    Observed band size: 38 kDa
    Additional bands at: 105 kDa, 65 kDa. We are unsure as to the identity of these extra bands.

  • All lanes : Anti-Torsin A/DYT1 antibody (ab34540) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Liver (Mouse) Tissue Lysate - normal tissue
    Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 5 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 38 kDa
    Observed band size: 38 kDa
    Additional bands at: 65 kDa, 65 kDa. We are unsure as to the identity of these extra bands.

  • ICC/IF image of ab34540 stained SHSY5Y cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab34540, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • IHC-FoFr image of Torsin A/DYT1 (ab34540) on Rat Cerebral Cortex section. The sections used came from animals perfused fixed with Paraformaldehyde 4%, in phosphate buffer 0.2M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.

    See Abreview

References

This product has been referenced in:

  • Bonsi P  et al. RGS9-2 rescues dopamine D2 receptor levels and signaling in DYT1 dystonia mouse models. EMBO Mol Med 11:N/A (2019). Read more (PubMed: 30552094) »
  • Beauvais G  et al. Efficient RNA interference-based knockdown of mutant torsinA reveals reversibility of PERK-eIF2a pathway dysregulation in DYT1 transgenic rats in vivo. Brain Res 1706:24-31 (2019). Read more (PubMed: 30366018) »
See all 25 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Thank you for confirming those details.

I have now arranged for the antibody ab125437 to be sent to you as well as the lysate ab29467. These are currently on the order number 1121023however they may be moved from this order through consolidation. If you would like to know the progress of these products please quote"xxxxxxxxxxxx; to our customer service representatives.

I hope this has been of help. Please do let us know how the customer gets on with these new antibodies.

Read More

Question

The customer has come back and said;

Thank you for sending us the replacement antibody for Anti-Torsin A (ab34540). Unfortunately, this replacement gives us the same results as the first antibody that you sent us (the incorrect size band on Western Blot). We have spent a lot of time validating this antibody and feel that, for whatever reason, this antibody is not suitable to use.

We would therefore like to test another TOR1A antibody. We have looked at the available antibodies for TorsinA on your website and have decided that Anti-Torsin A antibody (ab125437) may be appropriate. However, we feel that due to the initial antibody picking up the completely wrong protein, we are hoping that you will provide this for free for us to trial.

We also would like a positive and (if you are able to get one) a negative control. The positive can either be cells overexpressing Tor1a or the actual recombinant protein (ab114564). We do need large volumes of these controls (small aliquots) as only a small amount is needed to verify the antibody. We have also shown our results to a protein chemist with expertise in antibody validation and he believes that our optimisation steps have been thorough and that we need to test another antibody with positive and negative controls. I hope that you understand our position and that you will be willing to help us.

We would be very interested to know if Abcam will test this antibody by Mass Spectrometry to verify that it does indeed pick up Tor1A. We really would like to see the results.

Could you please advise?
Thanks for your help!

Kind regards,

Read More
Answer

Thank you for supporting this case.

I have had a further look into the case and I would like to offer the customer the anti-Torsin A antibody they have requested (ab125437), free of charge. Please let me know if they would still like for this to be arranged.

I may be able to also offer the recombinant protein, ab114564, however I would suggest that a more appropriate positive control would be to use the lysate used with the antibody previously, human fetal brain lysate, as this would allow the customer to observe the background as well as detect the endogenously expressed human protein. I could therefore provide a lysate such as https://www.abcam.com/Brain-Human-Tissue-Lysate-fetal-normal-tissue-ab29467.htmlif they would like.

I look forward to receiving your reply.

Read More

Question

The customer has come back and said;

Thank you for sending us the replacement antibody for Anti-Torsin A (ab34540). Unfortunately, this replacement gives us the same results as the first antibody that you sent us (the incorrect size band on Western Blot). We have spent a lot of time validating this antibody and feel that, for whatever reason, this antibody is not suitable to use.

We would therefore like to test another TOR1A antibody. We have looked at the available antibodies for TorsinA on your website and have decided that Anti-Torsin A antibody (ab125437) may be appropriate. However, we feel that due to the initial antibody picking up the completely wrong protein, we are hoping that you will provide this for free for us to trial.

We also would like a positive and (if you are able to get one) a negative control. The positive can either be cells overexpressing Tor1a or the actual recombinant protein (ab114564). We do need large volumes of these controls (small aliquots) as only a small amount is needed to verify the antibody. We have also shown our results to a protein chemist with expertise in antibody validation and he believes that our optimisation steps have been thorough and that we need to test another antibody with positive and negative controls. I hope that you understand our position and that you will be willing to help us.

We would be very interested to know if Abcam will test this antibody by Mass Spectrometry to verify that it does indeed pick up Tor1A. We really would like to see the results.

Could you please advise?
Thanks for your help!

Kind regards,

Read More
Answer

Thank you for updating us of the progress made with the Anti-Torsin A antibody (ab34540).

xxxxx is currently away from the office but I will try my best to help the customer resolve this case.

In order to understand the problems encountered with this antibody I would like to collect a few more details from you. It would be very helpful if you could ill in the questionnaire I have attached to this email. This will allow me to understand any protocol changes made since first using the antibody. If you could also include the images obtained with the two different lots of ab34540 that would be very helpful indeed.

I am sorry for the inconvenience this problem has caused you. I look forward to receiving your reply.

Read More

Answer

Thank you for kindly confirming these details. I am sorry this vial of antibody has not worked for this customer.

As requested, we are pleased to arrange a free of charge replacement. I can confirm that 1 vial of ab66587 has been added to your order number PO-14509. This has Abcam order reference 1107820.

I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

Read More

Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will provide us with vital information for our monitoring of product quality

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad vial.

I apologize for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation if the items has been purchased within the 6 month guarantee period. In order to arrange this, I would appreciate if you could confirm the order number and date of purchase.

In addition, I can suggest you may like to consider the following suggestions as a check for future experiments:

1. I can suggest to try blocking in BSA rather than milk. changing blocking agent can sometimes help to improve the results.

2. Is the current vial of secondary antibody working well with other primary antibodies. I can suggest to try including a no primary control which will help to assess if the secondary antibody is binding non specifically.

3. Heating to 95oC to reduce and denature can sometimes cause aggregation of protein. I can suggest to try 70oC instead for 10 minutes.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More

Answer

Thank you for your enquiry.

The exact immunogen sequence for ab34540 is as follows:

CKTVFTKLDYYLDD

I am sorry to confirm this has not been specifically tested for cross reactivity to TOR1B. However, I have conducted a BLAST search of the immunogen sequence, and this is not indicating any significant homology of the immunogen to other Torsin family members, including TORB1.Therefore I would suggest this antibody would not detect other Torsin family members although we have no specificdata from the lab to confirm this.

The customer may want to take a closer look:
http://blast.ncbi.nlm.nih.gov/Blast.cgi

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain - Cerebral Cortex)
Specification
Brain - Cerebral Cortex
Fixative
Paraformaldehyde

Dr. Sophie Pezet

Verified customer

Submitted Sep 07 2010

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293 cells transfected with human Torsin A)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
50 µg
Specification
HEK 293 cells transfected with human Torsin A
Blocking step
LI-COR blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jun 12 2009

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