概述

  • 产品名称
    MCF7全细胞裂解物
    参阅全部 MCF7 细胞裂解液
  • 常规说明
    Source - 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.

  • 经测试应用
    适用于: WBmore details

性能

  • 存放说明
    Store at -80°C. Avoid freeze / thaw cycle.
  • 存储溶液
    Constituents: 5% Bromophenol blue, 2% Beta mercaptoethanol, 4.4% SDS, 7.9% Tris HCl, 20% Glycerol
  • Concentration information loading...
  • 裂解物说明
    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.
  • 研究领域
  • 背景
    MCF-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.

应用

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.

应用 Ab评论 说明
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.

文献

This product has been referenced in:
  • Park YM  et al. Time since menopause and skeletal muscle estrogen receptors, PGC-1a, and AMPK. Menopause 24:815-823 (2017). Read more (PubMed: 28195989) »
  • 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 all 2 Publications for this product

客户评价及客户问答

1-5 of 5 Q&A

Question
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 am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement as well as with the positive control lysate as goodwill. The order number is xxxxx. 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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Question
Answer

Thank you for contacting us.

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 888-772-2226 and mention this order number.

I hope this information helps. Please contact us with any other questions.

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Answer

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 results, please let me know and I will be happy to offer you either replacements with alternative antibodies or credits on your original purchase. Please let me know how you would like to proceed and I will be happy to assist you further.

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Answer

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 tested the lysate wtih beta-actin to confirm that it is not degraded. Please do not hesitate to contact me if you have any additional questions or concerns.

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Answer

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, ab29537. This is also an MCF7 whole cell lysate, in a loading buffer with a different formulation than that of ab3871: Click here (or use the following: https://www.abcam.com/index.html?datasheet=29537). Please let me know if you would like to try this. If so, I will send it free-of-charge.

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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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