HT-29 whole cell lysate (ab3952)
Overview
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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.
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Tested applications
Suitable for: WBmore details
Properties
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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
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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 |
|---|
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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.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab3952 has not yet been referenced specifically in any publications.