Overview

  • Product nameMCF7 whole cell lysate
    See all MCF7 lysates
  • General notesSource - MCF7 cells (human) from ATCC (cat# HTB-22).


    MCF7 whole cell lysate was prepared by homogenization in modified RIPA buffer (150 mM NaCl, 50 mM Tris-HCl, pH 7.4, 1% Triton X-100, 1 mM EDTA, 1 mM PMSF,5 µg/ml Aprotinin, 1 µg/ml Pepstatin-A, 2 µg/ml Leupeptin, 1 mM Na3VO4, 1 mM NaF). Debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The lysate was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecyl sulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.

  • Tested applicationsSuitable for: WBmore details

Properties

  • Storage instructionsStore at -80°C. Avoid freeze / thaw cycle.
  • Storage bufferConstituents: 5% Bromophenol blue, 2% Beta mercaptoethanol, 4.4% SDS, 7.9% Tris HCl, 20% Glycerol
  • Concentration information loading...
  • Lysate notesMCF7 whole cell lysate was prepared by homogenization in modified RIPA buffer (150 mM NaCl, 50 mM Tris-HCl, pH 7.4, 1% Triton X-100, 1 mM EDTA, 1 mM PMSF,5 µg/ml Aprotinin, 1 µg/ml Pepstatin-A, 2 µg/ml Leupeptin, 1 mM Na3VO4, 1 mM NaF). Debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The lysate was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecyl sulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.
  • Research areas
  • BackgroundMCF-7 cells are a well-characterized estrogen receptor (ER) positive control cell line (cells are positive for cytoplasmic estrogen receptors) and therefore are a useful in vitro model of breast cancer to study the role of estrogen in breast cancer. Cells are also positive for cytokeratin and negative for desmin , endothelin , GFAP, neurofilament, vimentin.

Applications

Our Abpromise guarantee covers the use of ab3871 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. MCF7 whole cell lysate is ready to load on SDS-PAGE for Western blotting. We recommend loading 20 µg per lane for mini gels.

References for MCF7 whole cell lysate (ab3871)

This product has been referenced in:
  • Lau JY  et al. Differential expression of peroxisome proliferator activated receptor gamma and cyclin D1 does not affect proliferation of asthma- and non-asthma-derived airway smooth muscle cells. Respirology 15:303-12 (2010). WB . Read more (PubMed: 20070588) »

See 1 Publication for this product

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has cau...

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I am sorry to hear that you are having some trouble with the antibody. I am sending you a free positive control lysate of MCF7 cells, ab3871. To check the status of this order, please call Customer Service at 888-...

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Thank you for contacting us.

As we discussed on the phone, you may be able to improve your WB results by switching from milk to BSA and by using a higher concentration of primary antibody (1:500 for example). If you are unable to improve your...

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Thank you for contacting Abcam regarding ab58803.

I am sorry that you have been experiencing difficulties with this antibody in WB. As you discussed with my colleague on the phone, I look forward to hearing back from you after you have teste...

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Thank you for the clarification. That is an unusual result, if the DDR1 in the MCF7 is intact. We have not received other complaints about the lysate. We do not have a different lot of ab3871, so I suggest trying a similar product, ab2953...

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