A549 whole cell lysate (ab7910)
Overview
-
Product name
A549 whole cell lysate -
General notes
Cell line: A549 (Human lung adenocarcinoma epithelial).
Growth media: DMEM and 10% NCS (Newborn calf serum).A549 cell lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 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
A549 cell lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 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 minutes 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
A-549 cells are used as an in vitro model for a type II pulmonary epithelial cell model for drug metabolism. Cells are cytokeratin positive by immunoperoxidase staining. This line was initiated in 1972 by D.J. Giard, et al. through explant culture of lung carcinomatous tissue from a 58-year-old Caucasian male (PubMed 4357758). Further studies by M. Lieber, et al. revealed that A549 cells could synthesize lecithin with a high percentage of desaturated fatty acids utilizing the cytidine diphosphocholine pathway (PubMed 175022).
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab7910 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 concentration.
A549 cell lysate is ready to load on SDS-PAGE for Western blotting, 20 µg per lane is recommended for mini gel. |
| Notes |
|---|
|
WB
Use at an assay dependent concentration. A549 cell lysate is ready to load on SDS-PAGE for Western blotting, 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
-
SDS download
-
Datasheet download
References (1)
ab7910 has been referenced in 1 publication.
- Yougbare I et al. NCS 613, a Potent PDE4 Inhibitor, Displays Anti-Inflammatory and Anti-Proliferative Properties on A549 Lung Epithelial Cells and Human Lung Adenocarcinoma Explants. Front Pharmacol 11:1266 (2020). PubMed: 32973507