Nuclear Extraction Kit

$256.00
NE-50

RayBio® Nuclear Extraction Kit. This kit can process 50 extractions of 10

Product Features

  • Process 50 extractions of 107 cells each
  • Less than 10% contamination between nuclear and cytoplasm fractions
  • 2 hour processing time
  • Produces ~1 mg with with 2 mg/ml non-denature cytoplasm protein and ~0.5 mg with 5 mg/ml non-denature nuclear protein

Species

Mammalian cells or fresh tissue

Application Notes

Kit Components
  • NE Reagents-I, -II, and -III
  • Protease Inhibitor Cocktail
  • Phosphatase Inhibitor Cocktail I
  • Phosphatase Inhibitor Cocktail II
Other Materials Required
  • 1X PBS Buffer
  • Precision pipettes to deliver 1 µl to 1 ml volumes
  • Adjustable 1-25 ml pipettes for reagent preparation
  • Tissue homogenizer for tissue samples
  • Refrigerated centrifuge with swing out rotor and refrigerated microcentrifuge
  • Other lab consumables
Protocol Outline
  1. Prepare all reagents and samples as instructed in the manual.
  2. Add appropriate amount of 1X NE Reagent-I for sample amount
  3. Culture Media: Disperse Cells
    Tissue Samples: Homogeize using dounce homogenizer or tissue grinder
  4. Incubate 15 minutes on ice
  5. Add appropriate amount of 1X NE Reagent-II for sample amount
  6. Incubate 2 minutes on ice
  7. Centrifuge tubes at 4°C, 14000 x g for 30 seconds in microcentrifuge.
  8. Transfer supernatant (cytoplasm fraction) into new tube. Store at -80°C
  9. Add appropriate amount of 1X NE Reagent-III for sample amount
  10. Vortex on highest setting for 10 seconds to suspend pelletes completly
  11. Incubate for 40 minutes on ice, vortexing for 10 seconds every ten minutes
  12. Centrifuge tubes at 4°C, 14000 x g for 10 minutes in microcentrifuge. Transfer supernatant (nuclear fraction) to a new tube and store at 0-80°

Storage/Stability

NE reagent-I, II, and III may be stored for up to 6 months at 2° to 8°C from the date of shipment. Protease Inhibitor and Phosphatase Set-I, Set-II should be stored at -20°C or -80°C (recommended at –80°C). Note: the kit can be used within one year if the whole kit is stored at -20°C. Avoid repeated freeze-thaw cycles.

Product Specifications

Size:
50 extractions
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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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