Overview

  • Product name
    HT29 whole cell lysate
  • General notes
    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 (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1%sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.

  • Tested applications
    Suitable for: WBmore details

Properties

  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Constituent: 5% Beta mercaptoethanol
  • Concentration information loading...
  • Purity
    Whole Cell Lysate
  • Lysate notes
    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.
  • Research areas
  • Background
    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.

Applications

Our Abpromise guarantee covers the use of ab3952 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 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.

References

ab3952 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Q&A

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

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