Most commonly used lysis buffers

RIPA 150 mM NaCl 1 mM PMSF 1 mM EDTA 5 µg/ml Aprotinin 5 µg/ml Leupeptin 1% Triton x-100 1% Sodium deoxycholate 0.1% SDS 50 mM Tris, pH 7.4 NP-40 NaCl 150 mM NP-40 1% Tris, pH 8.0 50 mM HEPES Hepes pH 7.4 50mM NaCl 150mM Glycerol 10% Triton X-100 1% KCl 5mM EDTA […]

Odyssey infrared imaging system

Direct infrared fluorescence detection on the Odyssey Infrared Imaging System provides the established standard for Western blot analysis that can’t be equaled with chemiluminescence and visible fluorescence. Infrared detection gives you the quantitative analysis and wide linear dynamic range that chemiluminescence cannot. Strong and weak bands on the same blot are accurately detected without the […]

Everything about western blots

Everything you always wanted to learn about western blots but were too afraid to ask is in this web site: http://www.westernblotting.org/ Western blot protocols, troubleshoting, definitions, buffers and theory all in a web site. What is biotin? how the stripping buffer works? how do I prepare the lysis buffer? In a very basic way, you’ll find your answers in this […]

Western blot protocol

A complete western blot protocol from Howel lab in University of California San Diego. Here is the link: http://cancer.ucsd.edu/howelllab/Western.html From how to obtain a cell lysate to how to prepare the gels, running the gels, membrane transfer, detection and stripping the membrane. Also all the reagents and buffers you need for your western blot. There is […]

6X Laemmli buffer

Laemmli is a sample buffer to use in western blot.  6X sample buffer is added to each protein sample and is boiled or heated for 5-10 minutes. Recipe to prepare 10 ml: – 1.2gr SDS (sodium dodecyl sulfate) – 6mg bromophenol blue – 4.7ml glycerol – 1.2ml Tris 0.5M pH6.8 – 2.1ml dwater warm it […]