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

    21-22 of 22 Abreviews or Q&A

    Question

    BATCH NUMBER 87266 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Non-specific band Hela cells: two bands, one is 80kD, other is 38kD. SiHa and CaSki cells: three bands, 80kD, 42kD, and 38kD. C33A cells (HPV negative): four bands: 80kD, 48kD, 42kD, 38kD and 35kD SAMPLE Cell extract PRIMARY ANTIBODY Manufacturer: Abcam Species: mouse monoclonal [(C1P5)] Diluent: PBS Dilution: 1: 1,000 (1ug/ml) and 1:300 (3ug/ml) Incubation time: 4 ?, overnight wash step: TBST washing, 3? 10 min DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Positive control: HeLa, SiHa, Caski Negative control: C33A ANTIBODY STORAGE CONDITIONS Store at -20?. Store at +4? for short term. SAMPLE PREPARATION Pro-PREPTM protein extraction solution, Protease inhibitors and Heating sample. AMOUNT OF PROTEIN LOADED 35ug ELECTROPHORESIS/GEL CONDITIONS NuPAGE Novex Bis-Tris Gels, 4-12% of the gel TRANSFER AND BLOCKING CONDITIONS NuPAGE transfer buffer, 1 hour and 20 min, 5% milk blocking buffer SECONDARY ANTIBODY Manufacturer:[a competitor] Species: goat anti-mouse lgG Diluent: PBS Dilution: 1: 1,000 Incubation time: 2 hours at room temperature wash step: TBST washing, 3? 10 min HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Antibody dilution ADDITIONAL NOTES As your datasheet, this protein(16E6)size is 57kD, but our customer thinks that is 19kD. Could you let me know the reason the size of 16E6 is 57kD.

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    Answer

    I apologise for the delay in helping you with the enquiry regarding ab70, I have just now received information from the source of this antibody. I can confirm that the customer is correct, the detected size for E6 protein is indeed 17kda, I have corrected the datasheet immediately and apologize for the confusion. I enclose below the references where this antibody has been used. I hope these will help the customer. Can he/she check that the cells were adequately transfected? Abdulkarim B et al. Antiviral agent Cidofovir restores p53 function and enhances the radiosensitivity in HPV-associated cancers. Oncogene 21:2334-46 (2002). PubMed: 11948417 Kobayashi A et al. Lymphoid follicles are generated in high-grade cervical dysplasia and have differing characteristics depending on HIV status. Am J Pathol 160:151-64 (2002). PubMed: 11786409 Banks L et al. Identification of human papillomavirus type 18 E6 polypeptide in cells derived from human cervical carcinomas. J Gen Virol 68 ( Pt 5):1351-9 (1987). PubMed: 3033140 Below are my recommendations: 1) I would recommend to use TBST to dilute the antibody, not PBS, using 0.1% Tween20 as this is very important for the stability of the antibody. 2) We do not have experience with the Pro-PREPTM extraction buffer and this may also be a problem. Please ask your customer to use a RIPA buffer and to use plenty of protease inhibitors (fresh). 3) How was the sample heated? Was it boiled in loading buffer? Did the loading buffer contain reducing agents? 4) How long did he block the membrane for? Please ask him to try a different blocking agent (5%BSA) and to try different blocking times (30min, 1hr, overnight). 5) please ask the customer to test the secondary antibody alone and check if it gives similar problems with other primary antibodies My main concern is that the cells have not been transfected adequately and that therefore the levels of E6 protein are very low. Thank you for passing my comments to the customer and your support with this matter, I hope this will help,

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    Answer

    Thank you for your enquiry regarding ab70. The antibody recognises the E6 oncoprotein which can be found in the cytoplasm and/or in the nucleus. A paper by Lucia G. Le Roux and Junona Moroianu (Journal of Virology, February 2003, p. 2330-2337, Vol. 77, No. 4) says " The E6 oncoprotein of high-risk human papillomavirus type 16 (HPV16) interacts with several nuclear transcription factors and coactivators in addition to cytoplasmic proteins, suggesting that nuclear import of HPV16 E6 plays a role in the cellular transformation process. In this study we have investigated the nuclear import pathways of HPV16 E6 in digitonin-permeabilized HeLa cells. We found that HPV16 E6 interacted with the karyopherin (Kap) 2 adapter and could enter the nucleus via a classical Kap 2ß1-mediated pathway." I cannot comment on the quality of the staining of the review as we have not received any images unfortunately. The review has now been removed as it was felt that there was not enough information to help future customers, especially without an image. The antibody should be stored at 4C for a short term (1-6 months) and ideally should be aliquoted and stored at -80C for longer term storage. I hope this information helps, please do not hesitate to contact us if you need any more advice or information,

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    21-22 of 22 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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