Instruct: Identify the main category of Biorxiv papers based on the titles and abstracts
Query: A case for glycerol as an acceptable additive to cryoEM samples Buffer composition and sample preparation guidelines for cryo-electron microscopy are geared toward maximizing imaging contrast and reducing electron beam-induced motion. These pursuits often involve the minimization or complete removal of additives that are commonly used to facilitate proper protein folding and minimize aggregation. Among these admonished additives is glycerol, a widely used osmolyte that aids protein stability. In this work, we show that inclusion of glycerol does not preclude high-resolution structure determination by cryoEM, as demonstrated by a [~]2.3 [A] reconstruction of mouse apoferritin ([~]500 kDa) and a [~]3.3 [A] reconstruction of rabbit muscle aldolase ([~]160 kDa) in presence of 20% v/v glycerol. While we found that generating thin ice that is amenable for high-resolution imaging requires long blot times, the addition of glycerol did not result in increased beam-induced motion nor an inability to pick particles. Overall, our findings indicate glycerol should not be discounted as a cryoEM sample buffer additive, particularly for large, fragile complexes that are prone to disassembly or aggregation upon its removal.
20
The categories of this paper are: pharmacology
28
The categories of this paper are: communication
26
The categories of this paper are: biochemistry
11
The categories of this paper are: cognition
23
The categories of this paper are: immunology
25
The categories of this paper are: bioengineering
6
The categories of this paper are: plant
30
The categories of this paper are: scientific
31
The categories of this paper are: paleontology
7
The categories of this paper are: ecology
13
The categories of this paper are: animal
15
The categories of this paper are: developmental
10
The categories of this paper are: and
5
The categories of this paper are: cell
32
The categories of this paper are: zoology
0
The categories of this paper are: genetics