HT-29 whole cell lysate (ab3952)

Submit a review Q&A (3)

製品の概要

  • 製品名

    HT-29 whole cell lysate

  • 特記事項

    Cell line: HT-29 (Human colorectal adenocarcinoma grade II).
    Growth media: McCoy’s 5a medium with 1.5 mM L-glutamine, and 10% FBS.

     

    HT 29 cell lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylene diamine tetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecyl sulfate, 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.

  • アプリケーション

    適用あり: WBmore details

製品の特性

  • Mycoplasma free

    Yes

  • 製品の状態

    Liquid

  • 保存方法

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

  • バッファー

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

  • ライゼート 備考

    HT 29 cell lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylene diamine tetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecyl sulfate, 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 (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1%sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.

  • 研究分野

  • 背景

    HT29 cells are human intestinal epithelial cells which produce the secretory component of Immunoglobulin A (IgA), and carcinoembryonic antigen (CEA). Cells are used for tumourigenicity studies.

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab3952の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB
Use at an assay dependent dilution.

HT 29 cell lysate is ready to load on SDS-PAGE for Western blotting, 10 - 20 µg per lane is recommended for mini gel.
特記事項
WB
Use at an assay dependent dilution.

HT 29 cell lysate is ready to load on SDS-PAGE for Western blotting, 10 - 20 µg per lane is recommended for mini gel.

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 (0)

ab3952 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。

ab3952 は論文での使用が確認できていません。

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1-3 of 3 Abreviews or Q&A

Question

Mit welcher Methode wurden die Zelllinien verifiziert?
zweiter Anruf, 26.10.12: Kunde muss bestaetigt haben, dass keineeinzelne Zelle laenger als 6 Monate in Kultur war.

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Answer

Ich habe folgende Informationen erhalten, die für die Lysate ab3952, ab7900 und ab7909 gelten:

"The cell lines were procured from a reputable source (ATCC) and the flasks and cryopreservation vials are carefully labeled to maintain stock.

The cells are grown only when a new lot is required, with backups of the original line to minimize the number of passages they go through before being processed as lysates.

In terms of QC, we run SDS-PAGE gels and stain them with coomassie. The lysates are also used internally to validate our antibodies.

It is usually 2-4 weeks to grow enough cells for a batch of lysate, depending on how quickly the cells expand. I had the technician double-check and the current batch of lysate for ab3952 was in culture for 3 weeks, for example. She was also able to find the passage information – she thawed passage 2 for this lots."


Ich hoffe, dies hilft Ihnen weiter. Ich habe in der Zwischenzeit eine Erinnerung an den Hersteller von ab29545 gesandt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
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Question

boiling sample for 10 min and loading into gel. have called about previous vials with same problem. compared to home prepared lysates, very poor results, this product creates just a smear with comassie staining.

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Answer

Thank you for your call last week and for letting us know about the trouble with ab3952. My colleague is out of the office, but I am following-up with some of her emails. It is my understanding that you would like a credit note for a vial of ab3952 on the order ***. Could you please confirm this information, and I will process the credit note promptly.

I look forward to hearing from you. Please let me know if there is anything else that we can do for you, and I'll be happy to help.

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Question

Would happen to have any information on any housekeeping proteins for the cell line HT-29

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Answer

Thank you for your enquiry. HT-29 cells are human colonic Adenocarcinoma cells. We offer various loading control antibodies that target house keeping genes for use by various applications. I would recommend that you navigate to www.abcam.com and hover over our "product browser" before selecting the desired loading control. You can then select the antibody you desire based on your intended applications. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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