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Explore chemical space for cryoEM and crystallization

The Oryx8 in combination with the SpectroLight 600 can identify promising conditions for structure determination by cryoEM (and crystallization).

For cryoEM, screens such as the DIXC screen comprising of detergents, ions from the extremes of the Hofmeister series, buffers and substrates can be used. Favorable conditions can then be optimized before setting up cryoEM grids.

Oryx and SpectroLight

Screening and optimization experiments (with cryoEM and/or crystallography in mind) can be dispensed by the Oryx8, then efficiently analyzed by the SpectroLight600.

Several hundred samples can be analyzed per day, before choosing the best to pass on for analysis by cryoEM.

Representative results for a membrane protein in a 96 well detergent screen:

DLS Screen example



SpectroLight 600: In-Plate Dynamic Light Scattering System to Analyze, Monitor and Image Sub l Droplets

    Size distribution determination non invasively by in-situ DLS fully automated imaging system

    Operates on standard plates to perform high throughput in sub-microliter volumes

    Characterization of biomolecules for further sample evaluation like NMR, CryoEM, SAXS or Crystallization

    Assess homogeneity aggregation and assembly state

    Optimize buffer conditions, e.g. by selection of a specific detergent for membrane proteins solubility

    Investigate protein interactions, e.g. oligomerization, aggregation, denaturation, receptor-ligand interactions

    No cleaning required

DLS results example


Multi-channel nanoliter dispensing system

    Microbatch-under-oil is suitable for crystallization and CryoEM

    Oil prevents liquid air interface on drop

    Laser can analyse 140 nL drop under oil

    Only 7.0 l of protein required for a 96-well experiment (70 nl protein per drop)

    Hanging and sitting-drop vapor diffusion

    Auto design and gradient optimization with up to 7 ingredients

Oryx dispensing microbatch experiment


Microbatch optimization software Laser analysing microbatch-under-oil drop



DIXC screen for Cryo-EM   

27/10/2017

A new screen for Cryo-EM

Douglas Instruments has designed a screen that aims to improve the behaviour of macromolecules for cryoEM experiments. The screen includes ions at both extremes of the Hofmeister series (i.e. both chaotropes and kosmotropes), organic and inorganic buffers, detergents, sugars and alcohols. In order to reduce the noise generated by the ingredients of the screen in cryoEM images, low molecular weight compounds based on elements near the top of the periodic table were chosen where possible.

DIXC Conditions:

Well

Conc

Chemical

pH

1 (A1)

1 M

Mg chloride

2 (A2)

1 M

Li chloride

3 (A3)

1 M

Na chloride

4 (A4)

1 M

K chloride

5 (A5)

1 M

NH4 chloride

6 (A6)

1 M

NH4 sulfate

7 (A7)

1 M

Li nitrate

8 (A8)

1 M

Na formate

4

9 (A9)

1 M

Na citrate

10 (A10)

1 M

Na tartrate

11 (A11)

1 M

Na, K phosphate

7

12 (A12)

1 M

Na, K phosphate

4.5

13 (B1)

1 M

K thiocyanate

14 (B2)

1 M

Urea

15 (B3)

1 M

Guanidinium chloride

16 (B4)

1 M

Na malate

3.5

17 (B5)

1 M

Na acetate

5

18 (B6)

1 M

Na succinate

5.5

19 (B7)

1 M

Na maleate

6

20 (B8)

1 M

Na carbonate

6.5

21 (B9)

1 M

TEA

7.5

22 (B10)

1 M

Tris

8

23 (B11)

1 M

Bicine

8.5

24 (B12)

1 M

Na borate

9

25 (C1)

0.5 M

Glucose

26 (C2)

0.5 M

Fructose

27 (C3)

0.5 M

Galactose

28 (C4)

0.5 CMC

Fluorinated octyl maltoside

29 (C5)

0.5 CMC

Tetraethylene Glycol Monooctyl Ether (T350)

30 (C6)

0.5 CMC

Pentaethylene glycol monododecyl ether

31 (C7)

0.5 CMC

N-Decanoyl-N-methylglucamine

32 (C8)

0.5 CMC

n-Dodecyl -D-maltoside

33 (C9)

50 %

MPD

34 (C10)

50 %

Methanol

35 (C11)

50 %

Enthanol

36 (C12)

50 %

Isopropanol

37 (D1)

50 %

Glycerol

38 (D2)

50 %

Ethylene glycol

39 (D3)

0.1 %

Ultra-low melting point agarose

40 (D4)

1 M

Resorcinol

7

41 (D5)

1 M

Gly-Gly-Gly

7

42 (D6)

1 M

Oxamic acid

7

43 (D7)

1 M

Barbituric acid

7

44 (D8)

1 M

Na Malonate

7

45 (D9)

1 M

Glycerol 2 phosphate

7

46 (D10)

100 %

User defined

47 (D11)

100 %

User defined

48 (D12)

100 %

Water

 

 


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