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  1. Link

    ht-29-whole-cell-lysate-ab3952.pdf

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Kits/ Lysates/ Other Lysates Whole Cell Lysates Human Other
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HT-29 whole cell lysate (ab3952)

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

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Overview

  • Product name

    HT-29 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 (0.045 M Tris-HCl pH 6.8, 10% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue), containing 0.05 M DTT.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Mycoplasma free

    Yes
  • Form

    Liquid
  • Storage instructions

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

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

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

The Abpromise guarantee

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.
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.

Protocols

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

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab3952? Please let us know so that we can cite the reference in this datasheet.

ab3952 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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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.

Read More

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

Read More

Abcam Scientific Support

Answered on 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.

Read More

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.

Read More

Abcam Scientific Support

Answered on Jul 10 2012

Question

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

Read More

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.

Read More

Abcam Scientific Support

Answered on Apr 06 2006

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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