Overview

  • Product name
    A431 whole cell lysate
    See all A431 lysates
  • General notes
    Cell line: A431 (Human epidermoid carcinoma).
    Growth media: DMEM and 10% FBS.


    A431 cell lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 µg/ml of aprotinin, 5 µg/ml of leupeptin). Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The cell lysate was boiled for 5 minutes in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.

  • Tested applications
    Suitable for: WBmore details

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Constituent: 5% Beta mercaptoethanol
  • Concentration information loading...
  • Purity
    Whole Cell Lysate
  • Lysate notes
    A431 cell lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 µg/ml of aprotinin, 5 µg/ml of leupeptin). Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The cell lysate was boiled for 5 minutes in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.
  • Research areas
  • Background
    A431 cells are a positive control for EGFR as they express high levels of EGFR. They also used for cell cycle studies and can be stimulated with EGF to stimulate EGFR and are therefore a very useful model to study EGFR downstream signalling cascades eg shc, src, MAPK.

Applications

Our Abpromise guarantee covers the use of ab7909 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
WB Use at an assay dependent concentration. Ready to load on SDS-PAGE for Western blotting, 20 µg per lane is recommended for mini gel.

References

This product has been referenced in:
  • Niittykoski M  et al. Immunohistochemical Characterization and Sensitivity to Human Adenovirus Serotypes 3, 5, and 11p of New Cell Lines Derived from Human Diffuse Grade II to IV Gliomas. Transl Oncol 10:772-779 (2017). Read more (PubMed: 28797937) »
  • Awais R  et al. p63 is required beside p53 for PERP-mediated apoptosis in uveal melanoma. Br J Cancer 115:983-992 (2016). WB . Read more (PubMed: 27584665) »
See all 2 Publications for this product

Customer reviews and Q&As

1-4 of 4 Q&A

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I have issued a free of charge control lysate (ab30257) as goodwill with the order number. 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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Answer

Thank you for taking the time to look into this and for your response.

We have not tested this in HepG2 cells and the positive control recommendations were based on the literature. I thank you for your recommendation with this and have removed HepG2 from the controls.

Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Answer

Thank you for contacting us. I was able to find the reference that you spoke of on the phone (ShanLX, HardyMP Endocrinology.1992 Sep;131(3):1107-14. (PMID:1505454,http://www.ncbi.nlm.nih.gov/pubmed/1505454). During my research for this question I was able to find a later paper in whichHepG2 lines are shown to haveGnRHreceptors,Pati D,Habibi HR.Endocrinology.1995 Jan;136(1):75-84.(PMID:7828560; ). A more recent paper, Jacobson HI, et.al.Adv Exp Med Biol.2008;617:477-84. (PMID: 18497072,http://www.ncbi.nlm.nih.gov/pubmed/18497072) use HepG2 culture with hCG as well. Is seems that the early information has been supplanted by this new evidence. Therefore I believe that the HepG2 cells should be a positive control for this product. If you have further concern I would recommend using A431 cell lysate (https://www.abcam.com/A431-Human-epithelial-carcinoma-cell-line-Whole-Cell-Lysate-ab7909.html) or H1299 cells. 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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Answer

Thank you for your enquiry. For immunohistochemistry paraffin-embedded sections, human breast carcinoma was used in as control. For Western Blot QC, lysates were prepared from A431 cells, untreated or calyculin A-treated. A431 cells are a human epithelial carcinoma cell line. Calyculin treatment greatly increased the expression of the 14-3-3 Binding Motif, phosphorylated (Ser) in a number of different proteins. Abcam lists this product ab7909, A431 (Human epithelial carcinoma cell line) Whole Cell Lysate, which could serve as a decent positive control for Western Blotting.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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