In:
Journal of Biological Methods, Journal of Biological Methods, Vol. 3, No. 4 ( 2016-08-13), p. e51-
Abstract:
Washing and lysing monolayer cells directly from cell culture plasticware is a commonly used method for protein extraction. We found that multiple protein bands were enriched in samples with low cell numbers from the 6-well plate cultures. These proteins contributed to the overestimation of cell proteins and led to the uneven protein loading in Western blotting analysis. In Coomassie blue stained SDS-PAGE gels, the main enriched protein band is about 69 kDa and it makes up 13.6% of total protein from 104 U251n cells. Analyzed by mass spectrometry, we identified two of the enriched proteins: bovine serum albumin and bovine serum transferrin. We further observed that serum proteins could be extracted from other cell culture plates, dishes and flasks even after washing the cells 3 times with PBS. A total of 2.3 mg of protein was collected from a single well of the 6-well plate. A trace amount of the protein band was still visible after washing the cells 5 times with PBS. Thus, serum proteins should be considered if extracting proteins from plasticware, especially for samples with low cell numbers.
Type of Medium:
Online Resource
ISSN:
2326-9901
DOI:
10.14440/jbm.2016.129
Language:
Unknown
Publisher:
Journal of Biological Methods
Publication Date:
2016
detail.hit.zdb_id:
2849231-6
