L5178Y TK+/-

L5178Y TK+/- Whole Cell Lysate  

Catalog No: GRL-HLC037

Size: 200 μg 

Cell Line: L5178Y TK+/- (Mouse lymphoma)

Growth Media:

Dulbecco’s modified Eagle’s medium+4mM L-glutamine+1.5g/L sodium bicarbonate+4.5g/L glucose +0.1% Pluronic 90% fetal bovine serum 10%

Cell Lysate Preparation:  

。Prepare cell lysate by homogenization in ice-cold modified RIPA Lysis Buffer. 

。After centrifugation, determine the protein concentration of the cell lysate. 

。Boil the lysate for 5 minutes in 1X SDS sample buffer. 

。The lysate is ready to load onto SDS-PAGE for Western blotting. (20~50 μg per lane is recommended for mini gel.) 

Reagents: 

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.

SDS sample buffer: 50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% beta-mercaptoethanol. 

Application:

L5178Y TK+/- cell lysate is ready to load on SDS-PAGE for Western blotting, 20 μg~50 μg per lane is recommended for mini gel. 

Storage: 

It is supplied as whole cell lysate at 100 μg at 2 mg/ml in 1 x SDS sample buffer containing 5% beta-mercaptoethanol. Store at -20°C for three months or at –70°C for one year.