The largest database of trusted experimental protocols

Mini protean tgx precast 7.5 gel

Manufactured by Bio-Rad

The Mini-PROTEAN TGX Precast 7.5% Gel is a laboratory equipment product designed for protein electrophoresis. It is a pre-cast polyacrylamide gel that can be used for the separation and analysis of proteins based on their molecular weight.

Automatically generated - may contain errors

2 protocols using mini protean tgx precast 7.5 gel

1

Endothelial Cell Surface Biotinylation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Transduced ECs were starved for 3 h with EBM-2 medium (Lonza) at 37 °C, 5% CO2 in a humidified atmosphere. Cells were surface labeled with the cell impermeable HaloTag PEG-Biotin Ligand 3 µM at 4 °C for 15 min, stimulated or not with VEGF-A165 30 ng/ml for 10 min at 37 °C. ECs were lysed in buffer containing 25 mM Tris–HCl pH 7.6, 100 mM NaCl, 1% Triton X-100, 5% glycerol, 0.5 mM EGTA, 2 mM MgCl2, 1 mM PMSF, 1 mM Na3VO4 and protease inhibitor cocktail. Cellular lysates were incubated for 20 min on wet ice, and then centrifuged at 15,000 × g, 20 min, at 4 °C. The total protein amount was determined using the bicinchoninic acid (BCA) protein assay reagent (Pierce). Equivalent amounts (1 mg) of protein were immunoprecipitated for 2 h at 4 °C on streptavidin-conjugated beads. Immunoprecipitates were washed three times with lysis buffer with or without detergent and then separated by SDS–PAGE with Mini PROTEAN TGX precast 7.5% gel (BIO-RAD). Proteins were then transferred to a Trans-Blot Turbo™ PVDF Transfer (BIO-RAD), probed with antibodies of interest and detected by enhanced chemiluminescence technique (PerkinElmer).
+ Open protocol
+ Expand
2

Protein Extraction and Analysis via SDS-PAGE

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in buffer containing 25 mM Tris–HCl, pH 7.4, 100 mM NaCl, 5% glycerol, 0.5 mM EGTA, 2 mM MgCl2, 1 mM PMSF, 1 mM Na3VO4, protease inhibitor cocktail (Sigma-Aldrich), and detergent 1% NP-40. Cellular lysates were incubated for 20 min on ice, and then centrifuged at 15,000 g, 20 min, at 4°C. The total protein amount was determined using the bicinchoninic acid protein assay reagent (Thermo Fisher Scientific). Specific amounts of protein were separated by SDS–PAGE with precast Bolt 4–12% Bis-Tris gel (Thermo Fisher Scientific) or Mini PROTEAN TGX precast 7.5% gel (Bio-Rad). Proteins were then transferred to a nitrocellulose membrane (Bio-Rad), probed with Abs of interest, and detected by enhanced chemiluminescence technique (PerkinElmer).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!