Lysis-Extraction-Art-230

Cell lysis and organelle analysis

These kits are optimized for stepwise separation, enrichment and extraction of proteins from different cell fractions, including cytoplasmic, membrane, nuclear, chromatin-bound, and cytoskeletal proteins in 1–3 hrs. Most fractions are compatible with protein assays and typical downstream applications.

Select the right subcellular fractionation kit for your mammalian tissues or cells

NE-PER Nuclear Protein Extraction Kit Mem-PER Plus Membrane Protein Extraction Kit Cell Surface Protein Isolation Kit Subcellular protein fractionation kits (cells or tissues) Synaptic Protein Isolation Kit
Compatible sample types Tissues and cultured mammalian cells Tissues and cultured mammalian cells Cultured mammalian cells Tissues or cultured mammalian cells Brain tissue and primary neurons
Fractions isolated Nucleus, cytoplasm Integral and membrane associated Surface membrane proteins Nucleus, cytoplasm, membrane, cytoskeletal, chromatin bound Synaptic Proteins
Sample processing time 2 hrs 1 hr <1 hr 2–3 hrs <1 hr
Mechanical disruption required? Yes, for tissue Yes, for tissue No Yes, for tissue Yes, for tissue
Amount of Sample processed 50 samples with 2 million cells (20 µL packed) 50 samples with 5 million cells or 25 samples of 20 to 40 mg tissue per kit Eight experiments, with four confluent T75 flasks 25 extractions of 200 mg tissue or 50 samples with 2 million cells (20 µL packed) 10 g tissue or 500 x 35 mm dishes of primary cultured neurons
Protein assay compatibility BCA (CER dilute 1:4), Coomassie™ Plus (CER dilute 1:4), Pierce 660, Detergent Compatible Bradford BCA, Pierce 660, Detergent Compatible Bradford Pierce 660 (if compatibility reagent is used) BCA (all fractions), Pierce 660 (except PEB); NEB also compatible with Coomassie™ Plus and Detergent Compatible Bradford BCA
Downstream compatibility Both: western blot, ELISA, EMSA, reporter assays, enzyme assays,  amine reactive labeling
CER only: RNA EMSA, kinase assays, and RT-PCR
IP, western blot, ELISA, amine reactive labeling Western blot, ELISA IP, western blot, ELISA, EMSA, reporter assays, enzyme assays,  amine reactive labeling Neurotransmitter release assays, enzyme assays immunoassays, chromatography,
electrophoresis
Protease or phosphatase inhibitors recommended? Yes, both Yes, both Yes, both Both Included in kit Yes, both
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Subcellular fractionation overview

Schematic overview of the subcellular fractionation procedure. Cellular compartments are sequentially extracted by incubating cells with cytoplasmic extraction buffer (CEB) followed by membrane extraction buffer (MEB) and nuclear extraction buffer (NEB). Adding micrococcal nuclease (MNase) to NEB extracts chromatin-bound proteins from the cell pellet before adding the pellet extraction buffer (PEB) to solubilize cytoskeletal proteins.

Total protein profile of cytoplasmic and nuclear extracts prepared from different mouse tissues. Swiss Webster mouse tissues (40 mg) were harvested, rinsed with PBS and lysed using the NE-PER Nuclear and Cytoplasmic Extraction Reagent Kit. Extracts were quantified using the Pierce 660nm Protein Assay Reagent (Part No. 22660). Values are the average of two separate isolations.