Thermo Fisher Scientific
NE-PER™ Nuclear and Cytoplasmic Extraction Reagents
78835
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Mouse cells were given to a specific treatment, trypsinized and washed two times with PBS. The cell pellets were first resuspended with CER1 and then CER2 reagent, incubated, centrifuged and the supernatant was collected as cytoplasmic fraction. The pellet was then resuspended in NER reagent, incubated and isolated nuclear fraction. Both fractions were resolved in SDS PAGE and immunoblotted with compartment-specific protein antibodies.
Protein purification and isolation
Mouse cell, centrifuge
Proper incubation with reagent is essential
After the preparation of cytoplasmic and nuclear protein fraction, these fractions were analyzed by immunoblotting with compartment-specific proteins. Analysis reveals the presence of p53 proteins only in the nuclear fraction, not in cytoplasmic fractions.
None
Gave a two fraction of cytoplasmic and nuclear protein with high purity
There is a chance of 10% contamination between two fractions
Easy to use a kit to prepare cytoplasmic and nuclear protein with high purity.