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  1. Link

    products/cell-lysates/a-431-whole-cell-lysate-ab7909.pdf

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A-431 whole cell lysate (ab7909)

  • Datasheet
  • SDS
Submit a review Q&A (4)References (2)

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Overview

  • Product name

    A-431 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 min in 1 x SDS sample buffer (0.045 M Tris-HCl pH 6.8, 10% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue), containing 0.05 M DTT.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Mycoplasma free

    Yes
  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Constituent: 100% SDS Sample Buffer
  • Concentration information loading...
  • 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

    • Kits/ Lysates/ Other
    • Lysates
    • Whole Cell Lysates
    • Human
    • Other
  • 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

The Abpromise guarantee

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.
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.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (2)

Publishing research using ab7909? Please let us know so that we can cite the reference in this datasheet.

ab7909 has been referenced in 2 publications.

  • 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). 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 . PubMed: 27584665

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-4 of 4 Abreviews or Q&A

Question

If you do not have ovary cell lysate at present, the human testis tissue lysate will do, too.(https://www.abcam.com/Testis-Human-Tissue-Lysate-adult-normal-tissue-ab30257.html). Thank  you very much!

Read More

Abcam community

Verified customer

Asked on Feb 21 2012

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.

Read More

Abcam Scientific Support

Answered on Feb 21 2012

Question

I have read the two papers you mentioned about, but I still do not think they are very convincing. First, the  GnRH receptor is different from  HCG receptor either in  function or in structure. Even the existence of  GnRH receptor in HepG2 cells does not mean there are HCG receptors in them, too. And, although there are some changes when HepG2 cells being cultured with hCG, there is no direct proof that the HCG receptor is definitely there. While in the paper I mentioned(    Shan LX, Hardy MP  Endocrinology. 1992 Sep;131(3):1107-14.  ( PMID:  1505454,  ),  the authors did the Northern blot analysis showing there was no HCGR mRNA in HepG2 cells. I think this is a more solid proof. I want to know whether your company has ever tested the Anti-HCGR antibody on HepG2 cells . Because in my experiment, the Western blot showed some bands in HepG2 lane, but not at  79KDa band weight which is showed in your protocol. I do not know whether it is caused by the lack of specifity of this antibody. If you have tested the antibody in HepG2 cells, I want to know whether the results are the same. If you have not tested it, I do not think it is a good idea to list the HepG2 cells in the positive control before more convincing results been published. Thank you very much! Best wishes,

Read More

Abcam community

Verified customer

Asked on Feb 17 2012

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.

Read More

Abcam Scientific Support

Answered on Feb 17 2012

Question

Phoned because of concern regarding the positive controls listed on the datasheet. According to published reference, HepG2 is a negative control.

Read More

Abcam community

Verified customer

Asked on Feb 16 2012

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.

Read More

Abcam Scientific Support

Answered on Feb 16 2012

Question

What would be a suitable positive control for this product 14-3-3 (phospho S) antibody (0014-33)?

Read More

Abcam community

Verified customer

Asked on Jun 01 2007

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.

Read More

Abcam Scientific Support

Answered on Jun 01 2007

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