BN-PAGE was performed using the NativePAGE Bis-Tris Gel System (Thermo Fisher Scientific). Briefly, cells expressing both reporters were harvested without TfR silencing, washed with HBS, and solubilized in NativePAGE sample buffer supplemented with 10% glycerol, 1% DDM (n-dodecyl-β-d -maltoside), 1× protease inhibitor cocktail, and 100 μg/ml DNaseI. The samples were incubated in the solubilization buffer on ice for 30 min and centrifuged (13,000 × g at 4°C, 1 h), and the resulting supernatants were either untreated or treated with 4 M urea to denature protein complexes. Samples were then fractionated on precast 4–16% BN gradient gels (Thermo Fisher Scientific). After electrophoresis, proteins were transferred to polyvinylidene membranes (Millipore) and detected by our standard immunoblotting protocol with anti-HA or anti-VSG221.
Bn gradient gels
Manufactured by Thermo Fisher Scientific
4–16% BN gradient gels are electrophoresis gels used for the separation and analysis of protein complexes. These gels feature a continuous gradient of 4% to 16% polyacrylamide concentration, which allows for the effective separation of a wide range of protein complex sizes.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using bn gradient gels
1
Native Protein Complex Analysis
2
Native Protein Fractionation by BN-PAGE
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BN-PAGE was performed using the NativePAGE Bis-Tris gel system (Thermo). Briefly, cells were washed three times with 1X PBS supplemented with protease inhibitor cocktail without EDTA (Roche) and solubilized in Native PAGE sample buffer supplemented with 10% glycerol, 1% n-dodecyl-b-d-maltoside, 1x protease inhibitor cocktail without EDTA (Roche), 100 μg/ml microccocal nuclease (NEB), and 1x microccocal nuclease buffer (NEB). Samples were incubated in solubilization buffer on ice for 30 min and centrifuged (13,000 g at 4 ˚C, 30 min). The resulting supernatants were fractionated on precast 4–16% BN gradient gels (Thermo).
3
Native-PAGE Analysis of Protein Complexes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BN-PAGE was performed using the NativePAGE Bis-Tris Gel System (Thermo Fisher Scientific, Waltham, MA). Briefly, cells were harvested, washed with HBS and solubilized in NativePAGE Sample Buffer supplemented with 10% glycerol, 1% DDM (n-dodecyl-β-D-maltoside), 1X protease inhibitor cocktail and 100 μg/ml DNaseI. The samples were incubated in the solubilization buffer on ice for 30 min, centrifuged (13000g at 4°C, 1 hr), and the resulting supernatants were either untreated or treated with 4M urea to denature protein complexes. Samples were then fractionated on precast 4–16% BN gradient gels (Thermo Fischer Scientific). After electrophoresis, proteins were transferred to PVDF membranes (Millipore Corp., Bedford, MA) and detected by our standard immunoblotting protocol with anti-TfR or anti-VSG221.
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
Revolutionizing how scientists
search and build protocols!