A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate (ab7910)

Product Name 

A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate

Product type 

Lysates

Description 

A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate

Tested applications 
(see key)

WB

Application notes 
(see key)

Recommended dilutions
A549 cell lysate is ready to load on SDS-PAGE for Western blotting, 20 µg per lane isrecommended for mini gel.

Research areas 

Kits/ Lysates/ Other >> Lysates >> Whole Cell Lysates >> Human >> Other

Relevance 

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).


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.

Purity 

Whole Cell Lysate

Storage buffer 

Preservative: None
Constituents: 5% Beta mercaptoethanol, 12.5% Glycerol, 1% SDS, 1% Triton-X-100, 1% Sodium Deoxycholate, 0.01% Bromophenol blue, 5µg/ml Leupeptin, 5µg/ml Aprotinin, 50mM Tris HCl, 150mM Sodium chloride, 1mM EDTA, 1mM PMSF

Material safety datasheet (MSDS) for this product:
Mercaptoethanol MSDS

Form 

It is supplied as whole cell lysate, in 1 x SDS sample buffer containing 5% b-mercaptoethanol.

Concentration 

2.000 mg/ml

Storage instructions 

Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

Notes 

Cell line: A549 (Human lung carcinoma)
Growth media: DMEM and 10% NCS (Newborn calf serum)

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