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T-47D whole cell lysate (ab14899)

Submit a review Q&A (2)References (1)

製品の概要

  • 製品名

    T-47D whole cell lysate

  • 特記事項

    Extracts have been quality control tested by Western blot and the Electrophoretic Mobility Shift Assay (EMSA). 

製品の特性

  • Mycoplasma free

    Yes

  • 製品の状態

    Liquid

  • 保存方法

    Shipped on dry ice. Upon delivery aliquot and store at -80ºC. Avoid freeze / thaw cycles.

  • バッファー

    pH: 7.90
    Constituents: 0.75% Potassium chloride, 0.009% PMSF, 0.01% Magnesium chloride, 0.008% DTT, 0.48% HEPES, 20% Glycerol
  • Concentration information loading...

  • 研究分野

  • 背景

    T47D cells carry receptors for a variety of steroids and calcitonin. They express mutant tumor suppressor protein p53 protein. Under normal culturing conditions, they express progesterone receptor constitutively and are responsive to estrogen. They are able to lose the estrogen receptor (ER) during long-term estrogen deprivation in vitro. Cells are therefore used as a model for studies of drug resistance to tamoxifen in patients with mutant p53 breast tumors.

プロトコール

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

参考文献 (1)

ab14899 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。

ab14899 は 1 報の論文で使用されています。

  • Haglund F  et al. Prolactin receptor in primary hyperparathyroidism - expression, functionality and clinical correlations. PLoS One 7:e36448 (2012). PubMed: 22606260

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Question

I was looking at the above product on your website. I noticed that a comment was on the page asking about DNA and one of your colleagues responded saying that there would be no DNA present in the lysate. Would this also be true of RNA? And does the lysate only contain organelles and other cellular components? Another similar product was the same product treated with IFN gamma, are these essentially the same product and does this treatment cause any difference within the cells, for example in terms of expression levels of RNA or any other internal functions?

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Answer

Regarding ab14899, the procedure for the preparation of the whole cell extract is not RNAase free so the RNA will be at least partially degraded. But there is no step to get rid of the RNA and since it’s more soluble than DNA it could be present in the extract. We have never tested our extract for the presence of RNA and therefore we can not guarantee that it will be RNA free.

Unfortunately, we haven’t tested the presence of organelles either.

We have two nuclear lysates: ab14896 and ab14897 (IFN gamma treated).

These nuclear extracts don't contain any cytoplasmic proteins, the nuclei are first isolated then lysed. IFN gamma will affect the expression of some proteins. For example, they activate the expression of inflammation-related proteins.

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Question

I am inquiring to find out if it is possible to extract DNA from the T-47D cell lysate.

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Answer

Thank you for your enquiry. With the protocol used, the DNA is pelleted and discarded along with cell debris during the extraction process, so there is no DNA present in this lysate. I hope this information helps. Please do not hesitate to contact us if you need anything further.

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