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Anti-Torsin A/DYT1 抗体 (ab34540)

Reviews (2)Q&A (6)References (34)
Immunocytochemistry/ Immunofluorescence - Anti-Torsin A/DYT1 antibody (ab34540)
  • Western blot - Anti-Torsin A/DYT1 antibody (ab34540)
  • Western blot - Anti-Torsin A/DYT1 antibody (ab34540)

Key features and details

  • Rabbit polyclonal to Torsin A/DYT1
  • Suitable for: IHC-FoFr, ICC/IF, WB
  • Reacts with: Mouse, Rat
  • Isotype: IgG

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-Torsin A/DYT1 antibody [EP2569Y] (ab76133)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-Torsin A/DYT1 antibody
    Torsin A/DYT1 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to Torsin A/DYT1

  • 由来種

    Rabbit

  • 特異性

    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.

  • アプリケーション

    適用あり: IHC-FoFr, ICC/IF, WBmore details

  • 種交差性

    交差種: Mouse, Rat
    交差が予測される動物種: Human

  • 免疫原

    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)

  • ポジティブ･コントロール

    • ICC/IF: Neuro2a cells

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab34540の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
IHC-FoFr (1)
Use at an assay dependent concentration.
ICC/IF
Use a concentration of 1 µg/ml.
WB (1)
Use a concentration of 1 µg/ml. Detects a band of approximately 38 kDa (predicted molecular weight: 38 kDa).
特記事項
IHC-FoFr
Use at an assay dependent concentration.
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).

ターゲット情報

画像

  • Immunocytochemistry/ Immunofluorescence - Anti-Torsin A/DYT1 antibody (ab34540)
    Immunocytochemistry/ Immunofluorescence - Anti-Torsin A/DYT1 antibody (ab34540)

    ab34540 staining Torsin A/DYT1 in Neuro2a cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab34540 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

    Also suitable in cells fixed with 100% methanol (5 min).

    Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

  • Western blot - Anti-Torsin A/DYT1 antibody (ab34540)
    Western blot - Anti-Torsin A/DYT1 antibody (ab34540)
    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.

  • Western blot - Anti-Torsin A/DYT1 antibody (ab34540)
    Western blot - Anti-Torsin A/DYT1 antibody (ab34540)
    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.

参考文献 (34)

ab34540 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab34540 は 34 報の論文で使用されています。

  • Sharma R & Hetzer MW Disulfide bond in SUN2 regulates dynamic remodeling of LINC complexes at the nuclear envelope. Life Sci Alliance 6:N/A (2023). PubMed: 37188462
  • Ponterio G  et al. Alpha-Synuclein is Involved in DYT1 Dystonia Striatal Synaptic Dysfunction. Mov Disord 37:949-961 (2022). PubMed: 35420219
  • Tassone A  et al. Vesicular Acetylcholine Transporter Alters Cholinergic Tone and Synaptic Plasticity in DYT1 Dystonia. Mov Disord 36:2768-2779 (2021). PubMed: 34173686
  • Yokoi F  et al. Characterization of the direct pathway in Dyt1 ?GAG heterozygous knock-in mice and dopamine receptor 1-expressing-cell-specific Dyt1 conditional knockout mice. Behav Brain Res 411:113381 (2021). PubMed: 34038798
  • Scuteri D  et al. Effect of Gabapentin in a Neuropathic Pain Model in Mice Overexpressing Human Wild-Type or Human Mutated Torsin A. Life (Basel) 11:N/A (2021). PubMed: 33445430
View all Publications for this product

レビューと Q&A

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1-8 of 8 Abreviews or Q&A

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Torsin A/DYT1 antibody

 Good
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain - Cerebral Cortex)
Specification
Brain - Cerebral Cortex
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Sophie Pezet

Verified customer

投稿 Sep 07 2010

Western blot abreview for Anti-Torsin A/DYT1 antibody

 Excellent
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Jun 12 2009

Question

Hi Karin,

Please see below the customers response;

I am so sorry for the delay, but I was trying the replacement Antibody for the last time using wet transfer technique, but unfortunately the result is same by giving us the wrong band size.

Thank you for accepting our request by offering us for free the antibody (ab125437), recombinant protein (ab114564) and Brain (Human) Tissue Lysate - fetal normal tissue (ab29467), with the hope this will solve our problem. As you know, I am a PhD student and in desperate need to complete your project, especially i am running out of time.

Again, thank you for this generous offer which we really appreciate it.

I let the customer know that he needs to choose between the protein and lysate, he responded;

Thank you for your clarification. If we have to select one of the positive controls, I will go with the Brain (Human) Tissue Lysate - fetal normal tissue (ab29467) as Abcam suggested.

Thanks for your help!

Kind regards,

Read More
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

Question

Thank you for your help, please see below the response from the customer;

Thank you for your concern. I tried last Tuesday another gel using some of your colleague’s steps and some of our experience steps. Unfortunately, there is no improvement in the gel.

Therefore, I would like to ask you a free of charge replacement of this antibody with different batch number, hoping this will help to solve the problem and obtain satisfactory results.

Again, thank you for your lovely concern, which I really appreciate it.

Please let me know if you require any further information!

Kind regards,

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

Question

Hi Abcam,

Please find below an email we have received from a customer;

It is nice to write for you again and I hope this email finds you well and healthy. I am contacting you about the Anti-Torsin A antibody (ab34540) that I have ordered from Abcam 2 months ago, seeking your advice about technical issue.

In fact, I have conducted my western blotting experiment for TOR1A protein, but there was no detection for TOR1A protein at the right position (˜ 38kDa). In the same gel, I another bright band was detected at ˜110 kDa. I know it has been mentioned in the datasheet of this Ab that there are extra bands in different positions, but the strange is the interesting protein did not detected.

I tried to solve this by increase the amount of DTT (ranged between 200mm to 1000mm) which may help for further denatures for this homohexamer protein by reduce protein folding, BUT without improvement.

I have attached pictures of some of these experiments. Also, I have attached another document showing the buffers that I used (in each experiment I used 2 different buffers, including Ripa & WCE buffers).

Many thanks for your usual concern and support, which I really appreciate them.

I hope that you will be able to help, I have attached the technical enquiry form as above.

Kind regards,

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

Question

Hi Abcam,

A customer has the following enquiry;

could you please confirm whether this antibody does not cross react with TOR1B and the other family members? If you do not have reports about the specificity of this antibody for Torsin A, can you please provide me with the epitope of this antibody?

Thank you for your help!

Kind regards,

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

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