概述

  • 产品名称

    Anti-HPV16 E6 + HPV18 E6抗体[C1P5]
    参阅全部 HPV16 E6 + HPV18 E6 一抗
  • 描述

    小鼠单克隆抗体[C1P5] to HPV16 E6 + HPV18 E6
  • 宿主

    Mouse
  • 经测试应用

    适用于: WB, IP, IHC-Pmore details
  • 种属反应性

    与反应: Human papillomavirus
  • 免疫原

    Gel-purified HPV-18 E6-beta galactosidase fusion protein.

  • 阳性对照

    • Human cervical cancer cells.
  • 常规说明

    Clinical relevance: Detection of HPV in cervical smears and biopsies. Uses: Analysis of E6 expression in cell transformation studies.

    The E6 proteins are known to form oligomers; PMID19917295, PMID17209544.

性能

应用

Our Abpromise guarantee covers the use of ab70 in the following tested applications.

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

应用 Ab评论 说明
WB Use at an assay dependent concentration. Detects a band of approximately 16.5 kDa.

16.5 kDa band is HPV-18, 17 kDa band is HPV-16.

The E6 proteins are known to form oligomers; PMID19917295, PMID17209544.

IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

靶标

  • 相关性

    Human Papilloma virus (HPV) is responsible for the development of type and tissue specific papilloma in the oral cavity and in the larynx. In the skin papilloma virus can cause different types of warts and in the ano-genital region it is associated with condyloma or carcinoma. HPV types 1,6 and 11 usually are associated with benign transformations of the tissues, while e.g. HPV types 16, 18 and 31 seem to be responsible for the development of some carcinoma like cervical carcinoma. The role of HPV in non-melanoic tumours of the skin is under discussion. Protein E6 is a Transcriptional transactivator, binding double stranded DNA. It has transforming activity inactivating, with E6-AP ubiquitin-protein ligase, the human TP53/p53 tumor suppressor protein by targeting it for degradation.
  • 数据库链接

    • 别名

      • E6 antibody
      • Human Papilloma Virus antibody
      • Human papillomavirus type 16 E6 antibody
      • Human papillomavirus type 18 E6 antibody
      • Protein E6 antibody
      see all

    图片

    • Formalin-fixed, paraffin-embedded human uterine cervix squamous cell carcinoma tissue stained for HPV16 E6 + HPV18 E6 using ab70 at 1/200 dilution in immunohistochemical analysis.

      Incubated with primary antibody for 1 hour at 25°C.

      Heat mediated antigen retrieval at pH 6.

      See Abreview

    • Subconfluent proliferating MCF-7 or T47D cells were transiently transfected overnight with empty pcDNA3 vector, with FLAG wild-type E6 or zinc finger defective mutant E6 (E6-(C66,136G)). The cells were washed, post-incubated for 24 hours to allow gene expression, and harvested for Western blotting to detect the E6 oncoprotein using ab70.

    文献

    This product has been referenced in:

    • Wang X  et al. Human papillomavirus type 16 E6 oncoprotein promotes proliferation and invasion of non-small cell lung cancer cells through Toll-like receptor 3 signaling pathway. J Med Virol N/A:N/A (2017). WB ; Human . Read more (PubMed: 28480962) »
    • Xi R  et al. HPV16 E6-E7 induces cancer stem-like cells phenotypes in esophageal squamous cell carcinoma through the activation of PI3K/Akt signaling pathway in vitro and in vivo. Oncotarget 7:57050-57065 (2016). WB . Read more (PubMed: 27489353) »
    See all 17 Publications for this product

    客户评价及客户问答

    11-20 of 22 Abreviews or Q&A

    Answer

    I am afraid we do not have the immunogen sequence information for this antibody. I would suggest the following publications as some extra information for your customer but otherwise I cannot help on this occasion.

    http://www.ncbi.nlm.nih.gov/sites/entrez/3033140">Banks et al. 1987

    http://www.ncbi.nlm.nih.gov/pubmed/3018129">Matlashewski et al. 1986

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    Answer

    Thank you for your patience.
    Unfortunately, no additional information was received from the inventor.
    I am very sorry about that.
    I'd be happy to offer you either a free of charge replacement with a different antibody to either the same or another target, a credit or a refund.
    Please let me know which option would be best for you and I will arrange it.
    I look forward to hear back and resolve this case for you.

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    Answer

    Thank you for contacting us.
    I just heard back from the supplying lab:
    Unfortunately, we don't have a WB image. We have attached a reference for this antibody in case that is of any help. We shall pass on your questions to the inventor and hope for a more helpful response.
    I am sorry that it is taking longer than expected to obtain the anwers to your questions.
    In the meantime, could you please let me know your order or PO number?
    If the product was purchased within the last 6 months, I can offer you either a free of charge replacement, credit or refund.
    I will keep you informed about the details which I will hopefully receive soon from the lab.
    Please let me know about your order and how you would like to proceed.

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    Answer

    Thank you very much for your interest in ab70 .

    To our knowledge, ab70 (Anti-HPV16 E6 + HPV18 E6 antibody [C1P5]) has not been tested in ICC/IF.

    Therefore, I can offer a discount off a future purchase if you buy ab70 now, test it in ICC/IF and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

    If you are interested in this offer, please follow these steps:

    1. Reply to this e-mail to let me know that you would like to proceed and test ab70 in ICC/IF. I will then send a discount code. This code must be issued before purchasing ab70 so please wait for my reply before ordering.

    2. Purchase ab70 either by phone, fax, or online (www.abcam.com).

    3. Test it in ICC/IF.

    4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

    5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.

    We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab70 turns out to be unsuitable for ICC/IF, you will still receive the discount on your next purchase after your Abreview has been submitted.

    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/collaborationdiscount.

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    Question

    Dear technical support team: This customer has purchased ab70 (Anti-HPV16 E6 + HPV18 E6 antibody [C1P5]) and has conducted the WB several times. The results show no signal in HPV16 E6; therefore this customer wants to ask for your help to modify her experiment step, could you please offer any suggestion to improve her wb result? I attached the wb image in this letter and her experiment step as follow: 1. Order details: Batch number: gr45058-1 Po:943411 Abcam product code: ab70 Antibody storage conditions (temperature/reconstitution etc) -20 2. Please describe the problem (high background, wrong band size, more bands, no band etc). I used the anti-16E6/18E6 antibody (Ab70) to check the recombinant protein of 16E6 or 18E6 at the same time. It seems it can detect the 18E6 protein but not the 16E6 protein. 3. On what material are you testing the antibody in WB? · Species: E.coli ( rosetta, constructed into pGEX4T-1 vector) or Caski cell lysate (Human) · What’s cell line or tissue Caski cell · Cell extract or Nuclear extract: cell extraction · Purified protein or Recombinant protein: Recombinant protein or cell lysate 3. The lysate How much protein was loaded: at least 35 ug/well or the E.coli pellet after IPTG induction (with SDS sample buffer to denature What lysis buffer was used: Pierce E. coli extraction buffer or cell lysis reagent What protease inhibitors were used: PIC(Protease Inhibitor Cocktail from Pierce) What loading buffer was used: 6 X SDS sample buffer Phosphatase inhibitors: No Did you heat the samples: temperature and time: 100℃ for 5-10 min 4. Electrophoresis/Gel conditions/ Transfer conditions Reducing or non reducing gel: SDS-PAGE, denature form Reducing agent:6 X SDS-PAGE sample buffer Gel percentage : 12.5% Transfer conditions: (Type of membrane, Protein transfer verified): PVDF, 400 mA transfer 2hr 5. Blocking conditions Buffer:PBST Blocking agent: milk, BSA, serum, what percentage: 5% silk milk in PBST Incubation time:4℃ overnight Incubation temperature: 4℃ 6. Primary Antibody Species:mouse Reacts against: anti-16E6/18E6 (Human papillomavirus E6 protein) · At what dilution(s) have you tested this antibody:1:1000 · What dilution buffer was used: PBS · Incubation time: 1hr · Incubation temperature: RT · What washing steps were done: PBST wash 3 times at RT 7. Secondary Antibody Species:sheep Reacts against:anti-mouse IgG At what dilution(s) have you tested this antibody: 1:1000 Incubation time: 1hr Wash steps: PBST wash 3 times at RT Fluorochrome or enzyme conjugate: peroxidase Do you know whether the problems you are experiencing come from the secondary? No 8. Detection method ECl, ECl+, other detection method:ECL 9. Did you apply positive and negative controls along with the samples? Please specify. Ans: I used the anti-16E6/18E6 antibody (Ab70) to check the recombinant protein of 16E6 or 18E6 at the same time(18E6 as positive control, pre-induction pellet as negation control). It seems can detect the 18E6 protein but can't detect the16E6 protein. I have already confirmed the sequence of 16E6 construct is correct. 10. Optimization attempts · How many times have you tried the Western? At least 3 times · Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): No · Do you obtain the same results every time e.g. are background bands always in the same place? yes · What steps have you altered? extend the incubation time to overnight at 4℃ Thanks for your kindly help. Best regards

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    Answer

    According to our records, ab70 was proving difficult to use in WB and we were in contact in order to help resolve the issue. Could you update me if you are able to get good results with this antibody? We wish you the best of luck with your research and look forward to a reply.

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    Question
    Answer

    So that you can compare your staining data to published results, here are a couple references in which ab70 has been used in IHC: Lawson JS et al. Koilocytes indicate a role for human papilloma virus in breast cancer. Br J Cancer 101:1351-6 (2009). IHC-P. PubMed: 19773762 Birgersdotter A et al. Inflammation and tissue repair markers distinguish the nodular sclerosis and mixed cellularity subtypes of classical Hodgkin's lymphoma. Br J Cancer 101:1393-401 (2009). IHC-P; Human. PubMed: 19773754 I hope this is helpful. After checking the references, please let me know if you still feel the vial of ab70 you received are not working as expected.

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    Question
    Answer

    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 for one vial of ab70. 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.  

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    Question
    Answer

    Thank you for your phone call.  You may find the following dilution calculator helpful: https://www.msu.edu/~nixonjos/more/dilution.html?stock=1&needed=0.0001&volume=1&total=0.1 Please let me know if you have any additional questions. 

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    Question

    Dear technical support team: This customer has purchased ab70 (Anti-HPV16 E6 + HPV18 E6 antibody [C1P5]) and has conducted the WB several times. The results show no signal in HPV16 E6; therefore this customer wants to ask for your help to modify her experiment step, could you please offer any suggestion to improve her wb result? I attached the wb image in this letter and her experiment step as follow:  1. Order details: Batch number: gr45058-1 Po:943411 Abcam product code: ab70 Antibody storage conditions (temperature/reconstitution etc) -20 2. Please describe the problem (high background, wrong band size, more bands, no band etc). I used the anti-16E6/18E6 antibody (Ab70) to check the recombinant protein of 16E6 or 18E6 at the same time. It seems it can detect the 18E6 protein but not the 16E6 protein.   3. On what material are you testing the antibody in WB? ·         Species: E.coli ( rosetta, constructed into pGEX4T-1 vector) or Caski cell lysate (Human) ·         What’s cell line or tissue Caski cell ·         Cell extract or Nuclear extract: cell extraction ·         Purified protein or Recombinant protein: Recombinant protein or cell lysate  3.  The lysate How much protein was loaded: at least 35 ug/well or the E.coli pellet after IPTG induction (with SDS sample buffer to denature What lysis buffer was used: Pierce E. coli extraction buffer or cell lysis reagent What protease inhibitors were used: PIC(Protease Inhibitor Cocktail from Pierce) What loading buffer was used: 6 X SDS sample buffer Phosphatase inhibitors: No Did you heat the samples: temperature and time: 100℃ for 5-10 min  4.  Electrophoresis/Gel conditions/ Transfer conditions Reducing or non reducing gel: SDS-PAGE, denature form Reducing agent:6 X SDS-PAGE sample buffer Gel percentage : 12.5% Transfer conditions: (Type of membrane, Protein transfer verified): PVDF, 400 mA transfer 2hr 5. Blocking conditions Buffer:PBST Blocking agent: milk, BSA, serum, what percentage: 5% silk milk in PBST Incubation time:4℃ overnight Incubation temperature: 4℃  6. Primary Antibody Species:mouse Reacts against: anti-16E6/18E6 (Human papillomavirus E6 protein) ·         At what dilution(s) have you tested this antibody:1:1000 ·         What dilution buffer was used: PBS ·         Incubation time: 1hr ·         Incubation temperature: RT ·         What washing steps were done: PBST wash 3 times at RT  7. Secondary Antibody Species:sheep Reacts against:anti-mouse IgG At what dilution(s) have you tested this antibody: 1:1000 Incubation time: 1hr Wash steps: PBST wash 3 times at RT Fluorochrome or enzyme conjugate: peroxidase Do you know whether the problems you are experiencing come from the secondary? No 8. Detection method ECl, ECl+, other detection method:ECL 9. Did you apply positive and negative controls along with the samples? Please specify. Ans: I used the anti-16E6/18E6 antibody (Ab70) to check the recombinant protein of 16E6 or 18E6 at the same time(18E6 as positive control, pre-induction pellet as negation control). It  seems can detect the 18E6 protein but can't detect the 16E6 protein. I have already confirmed the sequence of 16E6 construct is correct. 10. Optimization attempts ·         How many times have you tried the Western? At least 3 times ·         Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control):  No ·         Do you obtain the same results every time e.g. are background bands always in the same place? yes ·         What steps have you altered? extend the incubation time to overnight at 4℃   Thanks for your kindly help.   Best regards  

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    Answer

    Thank you for your enquiry regarding ab70 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody. Additionally, thank you for supplying an image, as this has been very beneficial in understanding your concerns. The protocol looks fine to me however I would like to make few comments/suggestions that might help to improve your results and that you could consider trying: As the antibody gave expected results with 18E6 protein so I can say the antibody is fine. However I would suggest optimizing the protocol for 16E6 protein. Could you try either high concentration of 16E6 lysates or low dilution e.g. 1/500 of ab70? If the results do not improve please do not hesitate to contact me; I will then send you a free of charge replacement vial of same antibody.  

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    Answer

    Thank you for contacting Abcam. I have attached the MSDS for all except ab500, ChIP kit. I am still waiting for the laboratory to get back to me on that one and I will send it on as soon as I get it. If there is anything else I can do, please let me know.

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    11-20 of 22 Abreviews or Q&A

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