Cytoplasmic & Nuclear Protein Enrichment Kit
Fractionate cytoplasmic & nuclear proteins from cultured mammalian cells
- Minimal cross contamination
- Distinct nuclear and cytoplasmic protein isolates
- Versatile
- For adherent and non-adherent cultured cells
- Scalable protocol for variable number of cells
- Time-saving kit
- Includes 3 buffers
- Protocol takes less than 90 minutes
- Compatible with downstream applications including Western blotting
Figure 1. Binding of HDAC-1 nuclear specific antibody to nuclear fraction after fraction. The nuclear specific antibody HDAC-1 demonstrates lack of cross contamination between nucleus and cytoplasm after fraction with the AMRESCO Cytoplasmic & Nuclear Protein Enrichment kit. (1) Cytoplasmic (15ug) and (2) nuclear (12.75ug) proteins isolated from K562 cells with AMRESCO’s Cytoplasmic/Nuclear Fractionation Kit were resolved on a 12.5% Fluorescent Sprint NEXT GEL® (M318). Resolution of protein bands was viewed in the gel with a UV transilluminator (A) prior to transfer to PVDF membranes (E578). Immuno-blotting by the HDAC-1 (nuclear-specific) antibody (B) was detected using Visiglo™ HRP Chemiluminescent Substrate Kit (N218).
Figure 2. Distribution of NF-κB in unstimulated K562 cells. Proteins isolated from K562 cells using AMRESCO’s Cytoplasmic/Nuclear Fractionation Kit (Cytoplasmic and Nuclear) and Total Cell Lysis Buffer (Total). Proteins (10ug) were loaded onto a 12.5% ™ NEXT GEL® (M318), transferred to PVDF membrane (E578), immunoblotted with NF-κB p65 (Rel A) antibody and visualized with AMRESCO’s VisiGlo™ HRP Plus Chemiluminescent Substrate Kit (N219-Kit) .
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