Bcl-xL Δ27–82 and A142L (both without the membrane anchor) were produced as described previously.[7a (link), 15a (link)] The synthesis of 1 has been described.[7a (link)] Crystals of Bcl-xL Δ27–82 and 1 were obtained by mixing at a molar ratio of 1:1.3 then concentrating the sample to 10 mg/ml. Crystals were grown by the sitting drop method at room temperature in 0.2 M lithium nitrate, 20% (w/v) polyethylene glycol 3350. Prior to flash freezing in liquid N2, crystals were equilibrated into cryoprotectant consisting of reservoir solution plus increasing concentrations of ethylene glycol to a final concentration of 20%. Data collection and refinement methods are detailed in Table S1 .
Protocol detail
Structural Insights into Bcl-xL Complexes
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : Walter and Eliza Hall Institute of Medical Research
Other organizations : University of Wisconsin–Madison
Top 5 similar protocols
Protocol cited in 8 other protocols
Variable analysis
independent variables
- Bcl-xL Δ27–82 and A142L (both without the membrane anchor)
- Synthesis of 1
- Crystals of Bcl-xL Δ27–82 and 1 obtained by mixing at a molar ratio of 1:1.3 then concentrating the sample to 10 mg/ml
- Crystals grown by the sitting drop method at room temperature in 0.2 M lithium nitrate, 20% (w/v) polyethylene glycol 3350
- Crystals equilibrated into cryoprotectant consisting of reservoir solution plus increasing concentrations of ethylene glycol to a final concentration of 20%
- Molar ratio of Bcl-xL Δ27–82 and 1 (1:1.3)
- Concentration of the sample (10 mg/ml)
- Sitting drop method at room temperature
- Reservoir solution (0.2 M lithium nitrate, 20% (w/v) polyethylene glycol 3350)
- Cryoprotectant (reservoir solution plus increasing concentrations of ethylene glycol to a final concentration of 20%)
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!