Menu ccc

CYTOPLASMIC / NUCLEAR PROTEIN ENRICHMENT KIT

Cat.No. Size Price Quantity Subtotal Buy
M330-KIT 1 KIT 來電價

M330-KIT

Products Description

Overview

  • Generate distinct nuclear and cytoplasmic fractions from cultured cells in 90 minutes
  • For adherent or non-adherent cultured cells
  • Compatible with downstream Western blotting applications
 

Detailed Description

This kit isolates proteins from both the cytoplasmic and nuclear fractions of cultured cells. The cytoplasm and the nucleus are separated from one another by simple centrifugation steps following lysis in a non-ionic detergent. The entire procedure can be performed in less than 1.5 hours. Kit includes Fractionation Buffer 1, 1 ml, Fractionation Buffer 2, 1.5 ml, Fractionation Buffer 3, 30 ml, and Nonidet® P-40 Substitute, 0.1 ml. Contains sufficient material for 20 extractions of 1 x 106 cells. 

Resuspend cell pellet in a cytoplasmic lysis buffer for 15 minutes. Add detergent and pellet the nuclei. Transfer the supernatant containing the cytoplasmic fraction and further isolate the proteins with cytoplasmic extraction buffer. After the nuclei have been washed, the pellet is resuspended in nuclei lysis buffer for 15 minutes. The nuclei debris is pelleted and the supernatant contains the nuclei protein fraction.

 

Click the Information tab to learn more about the Cytoplasmic/Nuclear Protein Enrichment Kit

Information

Detection of β-tubulin and Histone H2B in cytoplasmic and nuclear lysates from K562 cells



Cytoplasmic (lanes A1, A3, B1, B3) and nuclear (lanes A2, A4, B2, B4) proteins were isolated using AMRESCO’s Cytoplasmic/Nuclear Protein Enrichment Kit (M330). The proteins were separated on a 12.5% Fluorescent SPRINT NEXT GEL® for 30 minutes at 300V and then visualized by UV transillumination (lanes A1, A2, B1, B2). The proteins were transferred to PVDF and then immunoblotted for β-tubulin (lanes A3, A4) and Histone H2B (lanes B3, B4) using RapidBlockTM Solution (M325) as the blocking solution and antibody diluent. The blot was developed using VisiGlo PlusTM HRP Chemiluminescent Substrate Kit (N219).


Related Products

  • Protease Inhibitor Cocktail 100X, General Use (M221)
  • Protease Inhibitor Cocktail 100X, General Use with EDTA (M222)
  • Protease Inhibitor Cocktail Mammalian (M250)
  • Phosphate Buffered Saline (PBS), 1X Sterile Solution (E504)
  • SeraFree™ Cryopreservation Media (RPMI) (N655)
  • SeraFree™ DMEM Cryopreservation Media (N676)
  • Trypsin 1:300 (0260)
  • DMSO, Ultra Pure Grade (N182)
  • Penicillin/Streptomycin, 100X (K952)