Thermo Scientific N-PER Neuronal Protein Extraction Reagent is a proprietary cell lysis reagent optimized for efficient extraction of proteins from all cellular compartments of neuronal tissue and primary cultured neurons.
Neuron cell lysates, such as brain tissue extracts, that are prepared using the N-PER Reagent are suitable for enzymatic activity assays (e.g., phosphatase, kinase, ATPase assays), immunoassays (e.g., Western blots, ELISAs, RIAs) and protein purification. The N-PER Lysis Reagent enables protein extraction to be completed in less than 30 minutes and requires mechanical disruption of tissue (e.g., Dounce Homogenization, Polytron). Typical neuronal protein yields are 70-90µg of protein per mg of brain tissue or 300µg of total protein from 106 primary neuronal cells.
- Optimized – efficient total protein extraction, including membrane proteins, from brain tissue or primary cultured neurons
- Gentle – preserves protein function without compromising yield
- Versatile – can be supplemented with protease inhibitors, reducing or chelating agents or required cofactors
- Compatible – extracts are suitable for use with total protein, enzymatic and immunological assays and protein purification methods
- Isolate functional proteins from neuronal tissues or primary neuronal cells
- Study enzymatic activity assays specific to neuronal tissue
- Generate lysates from neuronal tissues for biomarker analysis
Typical neuron structures include the cell body (soma), dendrites and an axon. The unique morphology and rich sphingolipid and cholesterol composition of neuron cell membranes creates challenges in preparing neuronal protein extracts. N-PER Neuronal Protein Extraction Reagent is specifically formulated to overcome the challenges of extracting functional native proteins from brain or other neuronal tissue and primary cultured neurons while increasing protein yield compared to other extraction methods.
The mild formulation of the N-PER Cell Lysis Reagent is ideal for downstream applications requiring active neuronal proteins. In addition, the N-PER Reagent is compatible with many applications including enzyme assays (e.g., phosphatase, kinase), immunoassays, various chromatography procedures and electrophoresis. Thermo Scientific Halt Protease Inhibitor Cocktail or Phosphatase Inhibitor Cocktail can be added just before use to prevent proteolysis or to offer additional protection from the high phosphatase activity normally present in brain tissue.
For each extraction, a ratio of 1g of tissue to 10mL of N-PER Reagent or 0.5-1mL N-PER Reagent per 10cm petri dish of primary cultured cells is recommended.
|Significantly higher yield of total neuronal proteins from mouse brain tissue. Homogenization of mouse brain neuronal tissue in Thermo Scientific N-PER Reagent resulted in greater overall protein yield than in our standard T-PER Tissue Protein Extraction Reagent and two varieties of phosphate-buffered saline (PBS).
|Better extraction of specific neuronal proteins. Thermo Scientific N-PER Reagent provided more consistent protein extraction of neuron-specific proteins than other lysis reagents tested. Yields with N-PER Reagent were greater for membrane-bound associated neuronal proteins (NMDAR2B, AMPA, PSD-95, synaptophysin, flotillin-1). N-PER Reagent also performed well for cytoplasmic proteins (tyrosine hydroxylase, MAPK). Interestingly, the reagent was less effective for the astrocyte-specific (i.e., non-neuronal) protein GFAP.
Review of cell lysis and protein extraction
Syn-PER Synaptic Protein Protein Extraction Reagent
Other cell lysis and fractionation reagents
Protease and Phosphatase Inhibitor Cocktails and Tablets
Pierce Tissue Strainers