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 […]

10X TBS (Tris Buffered Saline)

To prepare 1 liter of 10X TBS: 30 gr  Tris 88 gr NaCl 2 gr KCl mix everything in dwater until it’s disolved. Adjust pH at 7.5

Paraformaldehide 4%

To prepare 100ml of PFA 4% stock solution: – Add 86ml ddwater in an enlermeyer – Boil water in the microwave – Weigh 4g of PFA From now on in the fume hood – Transfer PFA in the erlenmeyer – Add a clean stirring bar – Add a drop of NaOH 10M – Stir it and keep […]

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 […]