A549 HCP ELISA Kit

Overview

Product name

A549 whole cell lysate

General notes

  • Cell line: A549 (Human lung epithelial adenocarcinoma).
  • 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 fluoride, 1% Triton X-100, sodium deoxycholic acid 1%, sodium deoxycholic acid 0.1% sodium dodecyl sulfate, aprotinin 5 µg/ml, leupeptin 5 µg/ml).

Cell fragments were removed by centrifugation. Protein concentration was determined with the 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 dodecyl sulfate, 0.01% bromophenol blue), which contained 0.05 M DTT.

Proven Applications

Suitable for: WB

Properties

Mycoplasma free: Yes

Way: Liquid

Storage instructions

Shipped at 4°C. At the time of aliquot delivery. Store at -80°C. Avoid the freeze/thaw cycle.

Storage buffer

  • pH: 7.20
  • Constituent: 100% SDS Sample Buffer

Concentration: 100µg to 2mg/mL

Lysing 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 fluoride, 1% Triton X-100, sodium deoxycholic acid 1%, sodium deoxycholic acid 0.1% sodium dodecyl sulfate, aprotinin 5 µg/ml, leupeptin 5 µg/ml). Cell fragments were removed by centrifugation. Protein concentration was determined with the 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 dodecyl sulfate, 0.01% bromophenol blue). containing 5% b-mercaptoethanol.

Bottom

A-549 host cell proteins 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 started in 1972 by D.J. Giard, et al. by explant culture of lung carcinomatous tissue from a 58-year-old Caucasian male (PubMed 4357758). Other studies by M. Lieber, et al. revealed that A549 cells could synthesize lecithin with a high percentage of unsaturated fatty acids using the cytidine phosphocholine pathway (PubMed 175022).

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