Western Blotting Sample Preparation Optimization
Protocols, optimization tips,
and more for Western Blot.
Selecting the best protein extraction kit
The first step to obtaining a clear, accurate Western blot is sample preparation. The critical step of sample preparation, the choice of protein extraction method, is a crucial one which ultimately makes the difference between a blank blot and a beautiful one. Boster supplies a range of lysis buffers and specialized extraction kits; use this guide to pick the right one for your task and start getting better results.
The classic ionic detergent, RIPA buffer is a powerful lysis agent. RIPA will solubilize all membranes, denature proteins, and release genomic DNA into the lysate. It is an excellent choice if you’re planning on denaturing your proteins for SDS-PAGE, but a poor one if you’re interested in studying protein interactions; RIPA buffer will break any protein interactions, rendering it useless for co-immunoprecipitations.
Mammal tissue/cell extraction kits
Boster offers optimized kits for protein extraction from mammalian cells and tissue samples. Boster Mammal Cell Protein Extraction Reagent is an optimized lysis buffer designed to enhance the solubilization of proteins without altering their functionality. This extraction kit is ideal for samples destined for Western blot or ELISA.
Bacterial cell extraction kit
The Boster bacterial cell extraction kit is designed to gently lyse bacterial cells with the aid of sonication. This kit contains a detergent-based lysis buffer, which must be dialyzed if you plan on using commassie blue stain or another detergent-sensitive assay. This kit is excellent for any type of bacterial cell lysis.
Membrane protein extraction kit
To extract membrane-bound proteins with their properties and interactions intact, use the Boster membrane extraction kit. This kit is designed to separate membrane-bound proteins from the plasma membrane and suspend them in a hydrophobic medium, preserving their enzymatic activity and conformation. This kit is ideal for studying membrane proteins that would otherwise be disrupted by traditional cell lysis techniques.
Cytoplasmic and nuclear extraction kit
This kit enables the rapid extraction of nuclear and cytoplasmic proteins while maintaining their native conformation and enzymatic activity. This kit is ideal for experiments requiring un-denatured and enzymatically active proteins, but may be excessively conservative for procedures like Western blot.
Mitochondrial extraction kit
Our most specialized kit, the mitochondrial extraction kit is capable of separating intact mitochondria from cells. The reagents are based on Boster’s unique technology that offers high yield and purity while maintaining the integrity of the mitochondria. This kit is ideal for any study that requires purified mitochondria, and offers a fast turnaround for rapid studies.
Keywords: Western blotting optimization, sample preparation, protein extraction kitsClick for more optimization tips