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Kits/ Lysates/ Other Lysates Whole Cell Lysates Human Other

HT-29全细胞裂解物(ab3952)

  • Datasheet
  • SDS
Submit a review Q&A (3)

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概述

  • 产品名称

    HT-29全细胞裂解物
  • 常规说明

    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.

  • 经测试应用

    适用于: WBmore details

性能

  • Mycoplasma free

    Yes
  • 形式

    Liquid
  • 存放说明

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

    pH: 7.20
    Constituent: 100% SDS Sample Buffer
  • Concentration information loading...
  • 裂解物说明

    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.
  • 研究领域

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

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab3952于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
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.
说明
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.

实验方案

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

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (0)

发表研究结果有使用 ab3952?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab3952 尚未被引用在任何文献中。

客户评价及客户问答

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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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Abcam community

Verified customer

Asked on Nov 05 2012

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!
https://www.abcam.com/abreviews

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Abcam Scientific Support

回复于 Nov 05 2012

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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Abcam community

Verified customer

Asked on Jul 10 2012

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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Abcam Scientific Support

回复于 Jul 10 2012

Question

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

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Abcam community

Verified customer

Asked on Apr 05 2006

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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Abcam Scientific Support

回复于 Apr 06 2006

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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