Anti-ERCC1 抗体 [8F1] (ab2356)

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製品の概要

  • 製品名

    Anti-ERCC1 antibody [8F1]
    ERCC1 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [8F1] to ERCC1

  • 由来種

    Mouse

  • アプリケーション

    適用あり: ICC/IF, WB, IP, IHC-P, IHC-Fr, Flow Cytmore details

  • 種交差性

    交差種: Rat, Human

  • 免疫原

    Recombinant full length protein corresponding to Human ERCC1.

  • ポジティブ･コントロール

    • Tonsil.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab2356 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 1 µg/ml.
WB 1/100 - 1/500. Predicted molecular weight: 33-36 kDa.
IP 1/100 - 1/500.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr 1/50 - 1/100. 1/50 - 1/100 in an ABC method. We suggest an incubation period of 30 minutes at room temperature.
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

ターゲット情報

  • 機能

    Structure-specific DNA repair endonuclease responsible for the 5'-incision during DNA repair.

  • 関連疾患

    Defects in ERCC1 are the cause of cerebro-oculo-facio-skeletal syndrome type 4 (COFS4) [MIM:610758]. COFS is a degenerative autosomal recessive disorder of prenatal onset affecting the brain, eye and spinal cord. After birth, it leads to brain atrophy, hypoplasia of the corpus callosum, hypotonia, cataracts, microcornea, optic atrophy, progressive joint contractures and growth failure. Facial dysmorphism is a constant feature. Abnormalities of the skull, eyes, limbs, heart and kidney also occur.

  • 配列類似性

    Belongs to the ERCC1/RAD10/SWI10 family.

  • 細胞内局在

    Nucleus.
  • Information by UniProt

  • 参照データベース

    • 別名

      • COFS 4 antibody
      • COFS4 antibody
      • DNA excision repair protein ERCC 1 antibody
      • DNA excision repair protein ERCC-1 antibody
      • DNA excision repair protein ERCC1 antibody
      • ERCC 1 antibody
      • ERCC1 antibody
      • ERCC1_HUMAN antibody
      • Excision repair cross complementation group 1 antibody
      • Excision repair cross complementing 1 antibody
      • Excision Repair Cross Complementing Rodent Repair Deficiency Complementation Group 1 antibody
      • Excision repair protein antibody
      • RAD 10 antibody
      • RAD10 antibody
      • UV 20 antibody
      • UV20 antibody
      see all

    画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERCC1 antibody [8F1] (ab2356)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERCC1 antibody [8F1] (ab2356)

      Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue with ab2356 labelling ERCC1.

    • Immunocytochemistry/ Immunofluorescence - Anti-ERCC1 antibody [8F1] (ab2356)
      Immunocytochemistry/ Immunofluorescence - Anti-ERCC1 antibody [8F1] (ab2356)
      ICC/IF image of ab2356 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2356, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Flow Cytometry - Anti-ERCC1 antibody [8F1] (ab2356)
      Flow Cytometry - Anti-ERCC1 antibody [8F1] (ab2356)
      Overlay histogram showing HeLa cells stained with ab2356 (red line). The cells were fixed with 100% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2356, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERCC1 antibody [8F1] (ab2356)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERCC1 antibody [8F1] (ab2356)

      Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil tissue sections with ab2356 labelling ERCC1.

    参考文献

    This product has been referenced in:

    See all 23 Publications for this product

    ab2356 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。

    レビューと Q&A

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    1-9 of 9 Abreviews or Q&A

    Question

    The researcher has used ab2356 in WB and gets no signal, when another anti ERCC1 antibody gives good results. He has tried 1:100-1:1000 with overnight incubation in TBST-5% milk (plus prior blocking in 5% milk). PO: PM07689 If the problem is due to the antibody he would like a replacement vial with ab43.

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    Answer

    I have received feedback from the source of the antibody ab2356 who informed me that the application "tested in WB" came from one of their customers and that they did not have information on the dilutions or blocking agents tested with this product. In light of this I have decided to remove the application from our datasheet to prevent further inconvenience to future users of this antibody and would like to apologize for this problem. I have arranged for a replacement vial of ab43 to be sent to you on Monday on order 131817, I hope this product will be to your satisfaction. I hope this unfortunate matter will not deter you from purchasing antibodies from us in the future and apologize once again for this problem, I wish you all the best with your research,

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    Question

    I try to do an immunoprecipitation with ERCC1 3H11. I see on your datasheet for this antibody that you advice to use it at a concentration of 2µg/mg of lysate. So I supposed that you have perhaps a protocol… If you have one, can you send it to me, please? Thank you.

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    Answer

    Thank you for contacting us.

    I have contacted the lab regarding the protocol used to do IP for ab43. Unfortunately, I have found out that we have no information regarding how this antibody functions in IP and therefore we cannot guarantee that this antibody will work in IP. I am sorry for this mistake on our part and I have removed this application from the datasheet.

    Since you bought this antibody whilst it was still guaranteed for use in IP, I propose one of two things:

    1. An alternative antibody ab2356 (https://www.abcam.com/ab2356) which is guaranteed to work in IP

    OR

    2. You could test ab43 in IP. If you agree to test this antibody for us and send us your results via an abreview, we will issue you with a discount code which will entitle you to a discount of the price of ab43 off your next order.

    Simply follow these easy steps below to apply for our AbTrial Program:

    1. Reply to this email, letting us know you are interested in testing this product.

    2. Our scientists will email you an inactive personal discount code for the value of the product.

    3. Test the product

    4. Submit your results, including your discount code in the additional notes section of your Abreview.

    5. Once the Abreview is submitted, the discount code will become active.

    6. Apply your discount code on your next order to receive that value off.

    Please let me know if you have any questions about this offer and I would be happy to help you further.

    The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

    If you would like a general IP protocol, please visit :
    https://www.abcam.com/index.html?pageconfig=resource&rid=11385

    I hope this information is helpful to you and I look forward to your reply regarding how you would like to proceed on this matter.

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    Question

    Inquiry: MY inquiry about Anti-ERCC1 antibody [8F1] (ab2356) by using Immunohistochemistry (Formalin/PFA-fixed paraffin ) i want to know the dilution factor if the size of the lot was 500 µg so how many µg will dilution in ml i want to know 500 µg will be sufficient to how many number of cases.

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    Answer

    Thank you for your enquiry and your interest in our products.

    I would suggest carrying out a preliminary experiment with different dilutions of this primary antibody (i.e. 1:100, 1:500 and 1:1000) and then adjust the dilution accordingly to your results obtained.

    Each antibody should be titrated to determine the optimal working dilutions. The required quantity of this antibody depends on many factors:

    1. dilution factor of the antibody

    2. expression levels of the target in the samples,

    3. how do you carry out the assay (in duplicates, triplicates or in quadruplicates)?

    4. presence of positive and negative control

    5. surface area of the tissue sections etc.

    If you need any further assistance in the future, please do not hesitate to contact me.

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    Question

    Inquiry: Please send me a reference for using the anti ERCC1 Ab (ab2356) in flow cytometry. Thanks,

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    Answer

    Thank you for your enquiry.

    I can confirm that any publications that we are aware of will be listed on the datasheet. We have a dedicated team who spend time reviewing the literature to find articles that have used our products.

    However, in many cases, regrettably we do not have access to all the articles mentioning our products, or subscriptions to all the journals that publish them.

    I'm sorry there are none to provide for flowcytometryat the momentfor ab2356.

    Should you have any further questions, please do not hesitate to contact us.


    IF CUSTOMER NOT HAPPY:


    I can confirm that any publications that we are aware of will be listed on the datasheets. We have a dedicated team who spend time reviewing the literature to find articles that have used our products. I understand you concerns, it is unfortunate that we are not always able to provide these. However, in some cases, regrettably we do not have access to all the articles mentioning our products, or subscriptions to all the journals that publish them.

    If there are any particular antibodies you would like some more information on, I would be pleased to investigate further with our sources in case there are some more publications to provide. In this case, I would appreciate if you would like to provide the product numbers and I can look into this for you.

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    Question

    feeback in survey and phone call

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    Answer

    Merci d'avoir pris le temps de m'expliquer la situation aujourd'hui au téléphone. Je suis très désolée que le ab2356 n'as pas non plus fonctionné.

    Voici le lien pour les huit anticorps contre le ERCC1 que nous avons actuellement dans notre catalogue qui sont testées en IHC-P et sur les coupes humaines. Veuillez s'il vous plaît me dire lequel vous intéresse le plus ou si vous avez des questions additionnelles sur ces anticorps.

    https://www.abcam.com/index.html?pageconfig=searchresults&search=ERCC1&pt=1&sk=react&sv=21&sn=Human&l=3&fViewMore=1

    J'ai contacté aussi le laboratoire et je vous tiens au courant dès que j'ai une réponse.

    Je vous souhaite une agréable soirée et je me réjouis d'avoir de vos nouvelles.

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    Question

    j'ai essayé avec une étape de permeabilisation avec du Triton 0,2% 20 min mais il n'y a toujours pas de marquage.

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    Answer

    Comme convenu j'ai mis en place l'envoi d'une unité de ab2356 en remplacement du ab59772. Le numéro de commande de remplacement gratuit de ce produit est ******. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

    N'hésitez pas à nous contacter lors d'une prochaine occasion.

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    Question

    pas de signal IHC-P avec ab59772.
    Placenta humain.détection HRP
    Démasquage antigene : pH 6.0 et pH 9.0 essayé. 20/30 minutes bain marie.
    lot GR26082-1.
    Etape de perméabilisation va être tentée. (protéine nucléaire)

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    Answer

    Merci de nous avoir contactés.

    Une perméabilisation d'environ 10 minutes avec 0.2% de Triton peut améliorer les résultats.

    Merci de m'informer si cette suggestion s'avère êtreutile et nous mettrons à jour la fiche technique.

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    Question

    On the FAQs for this product it mentions that it has been used unsuccessfully for western blots. It also states therefore that this has been removed frm the application notes for this product but this has not happened. My question therefore is has this antibody now been tested by western blot applications? I would like to use it for this but obviously do not want it if it still doesn't work! SAMPLE Rat cell extracts ANTIBODY STORAGE CONDITIONS N/A - have not bought it yet

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    Answer

    Thank you for contacting us. We had customer feedback in 2006 that this antibody does not work in western blot so we decided to remove this application. Since then we have sold hundreds of units of this antibody and again we have received common feedback from our customers that the antibody worked well in WB so we decided to publish the application again. I would suggest buying the antibody as normal for WB and submitting an Abreview. Please be also advised that use of this antibody in WB is fully covered by Abpromise guarantee. https://www.abcam.com/index.html?pageconfig=abpromise I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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    Question

    What is the concentration of this antibody? Was it purified from ascites or tissue culture supernatant?

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    Answer

    Thank you for your enquiry. The concentration is 100 ug/ml, and the antibody was purified from ascites. I hope this information helps. Please do not hesitate to contact us if you need anything further.

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