Protein extraction is the oft used method for isolation of proteins. A critical requirement of this step is efficient cell lysis and separation of organelles if specific proteins are to be extracted from these. The methods used for extraction depend on the type of tissue, samples and organelle needed to the separated. For more information visit: Synopsis.
Factors to consider in this step are type of cell disruption needed: Plant cells have cellular walls which are difficult to break, bacterial cell walls can be sonicated, tissues such as bones and others tough tissues benefit from freeze-fracturing. Multiple options for homogenization and cell disruption are available from OMNI International.
Secondly, use of detergents, protease inhibitors, protein stabilization agents are common to ensure protein function. This is necessary for applications such as ELISA and crystallography which require both function and structure of the proteins to be maintained. Furthermore, clinical samples such as blood, urine etc., contain inhibitors and proteases- hence choice of buffers and steps in the workflow should be made keeping the above parameters in mind.
An often used technique to purify protein in large abundance is to clone it in vectors and express it in suitable cells and cell lines.
Cell lysis solutions for these are:
- Bacterial Cell Lysis
- Insect Cell Lysis
- Yeast Cell Lysis
- Plant Cell Lysis
Solutions for culture mammalian cells and human tissues are: Protein Extraction Solutions
Additional resources on protein extraction
Technical guide and handbook
Popular products for Protein Extraction
Product | Types of Samples | Assay Compatibility | Compatible Applications |
---|---|---|---|
T-PER Tissue Protein Extraction Reagent Cat# 78510 |
Heart, liver, kidney, lung, spleen | BCA, Coomassie Plus, Pierce 660, Detergent Compatible Bradford | IP, Western blots, ELISA, EMSA, purification, kinase assays, activity assays & amine reactive labeling |
N-PER Neuronal Protein Extraction Reagent Cat# 87792 |
Brain tissue and primary neurons | BCA, Pierce 660, Detergent Compatible Bradford | IP, pulldowns, Western blots, ELISA, enzyme assays & amine reactive labeling |
M-PER Mammalian Protein Extraction Reagent Cat# 78501 |
Cultured Mammalian and primary cells | BCA, Coomassie Plus, Pierce 660, Detergent Compatible Bradford | IP, Western blots, ELISA, EMSA, reporter assays, purification, activity assays & amine reactive labeling |
RIPA Lysis Buffer Cat# 89900 |
Cultured Mammalian cells | BCA, Detergent Compatible Bradford (dilute 1:3) | Western blots, ELISA & purification |
IP Lysis Buffer Cat# 87787 |
Cultured Mammalian cells | BCA, Pierce Detergent Compatible Bradford | Western blots, ELISA, IP & reporter assays |
Additional Solutions for Protein Extraction
Protease Inhibitors | Catalogue Number |
---|---|
Pierce™ Protease Inhibitor Mini Tablets, EDTA-free | A32955 |
AEBSF Protease Inhibitor | 78431 |
E-64 Protease Inhibitor | 78434 |
Bestatin Protease Inhibitor | 78433 |
Aprotinin Protease Inhibitor | 78432 |
Halt™ Protease Inhibitor Cocktail, EDTA-Free (100X) | 78437 |
Pepstatin A Protease Inhibitor | 78436 |
Pierce Protease and Phosphatase Inhibitor Mini Tablets | A32959 |
Pierce Protease and Phosphatase Inhibitor Mini Tablets,EDTA-free | A32961 |
Detergents | Catalogue Number |
---|---|
Tween™ 80 Surfact-Amps™ Detergent Solution | 28328, 28329 28320 |
Triton™ X-100 Surfact-Amps™ Detergent Solution | 28314, 28332 |
NP-40 Surfact-Amps™ Detergent Solution | 28324, 85124, 85125 |
Brij™-35 Surfact-Amps™ Detergent Solution | 28316, 85117, 85118 |