C-Tube LCDM
For growing large protein crystals by dialysis and diffusion

Catalog #Product nameDescription

Price (w/o tax)

Regenerated cellulose membrane
MB2004-CRT810

C-Tube LCDM(RC)

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MWCO 6-8kD
6 setsCellulose ester membrane

MB2004-CRT820

C-Tube LCDM(CE)

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MWCO 1kD
6 sets

MB2004-CRT830

Polyethersulfone membrane
C-Tube LCDM(PES) MWCO 10kD

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6 sets◼ Crystallization cell◼
◼
◼
◼
◼

Size: 2.5mm O.D., 1.92mm I.D., 18mm length
Material: GlassSample volume: 30μL
Dialysis membrane: Regenerated cellulose membrane (RC) MWCO 6-8kD; cellulose ester
membrane (CE) MWCO 1kD; polyethersulfone membrane (PES) MWCO 10kD.
Pre-attached gel-tube.

◼ Other items：Gel soaking bag; crystallization bag; fine top tip; PTFE tubing tip; C-Cap.

Practical – Growing large Lysozyme crystal
1.
2.

Keep the gel-tube of C-Tube LCDM in 5% PEG 4000, 0.4 M NaCland 50 mM acetate buffer pH 4.5 to soak the solution.

Diffraction source

Aichi SR BL2S1

Wave length (Å)

1.12

Camera system

ADSC315r

Load 25 mg/ml lysozyme, 5% PEG 4000 and 50 mM acetate buffer
pH 4.5 into the C-Tube LCDM cell with the fine top tip.

Space group

P43 21 2

Unit-cell parameters(Å, °)

79.1 79.1 38.2
90.0 90.0 90.0

Average mosaicity (°)

0.07

Resolution range (Å)

39.54 – 1.39
(1.42 – 1.39)

Completeness (%)

99.9 (100)Rmerge0.028 (0.569)<I/σ(I) >41.4 (4.0)

3.

Reservoir solution #1: 5% PEG 4000, 0.4 M NaCl, 50 mM acetatebuffer pH 4.5. A small crystal starts growing (Fig. 1).

4.

Change the reservoir solution from #1 to #2: 5% PEG 4000, 0.7 M
NaCl, 50 mM acetate buffer pH 4.5. The crystal is growing larger(Fig. 2).

5.

Summary of X-ray diffraction
experiment on lysozyme crystal

Fig. 1

Fig. 2

Change the reservoir solution from #2 to #3: 25% PEG 4000, 0.7
M NaCl, 50 mM acetate buffer pH 4.5. The crystal is growinglarger. By using higher PEG solution, X-ray diffraction data ofhigher resolution is expected (Fig. 3).

Mn(I) half-set correlation
1.00 (0.923)
CC(1/2)
Fig. 3

TheMajor Features of C-Tube LCDM

Dialysis/diffusion method

Amount of protein sample

No protein loss during the crystallization experiment.
Osmotic pressure difference is small in C-Tube LCDM.

30μL of protein sample is required.
If Vm is 2.2Å3/D, one crystal of 1.2mm3 size can grow.

Crystallization condition
The crystallization condition which suppresses nucleation
probability should be chosen, considering the diffusion of protein
and precipitant molecules in the solution.
Please contact us for consultation and technical assistance.

Easy preparation

Long-term stability/space-saving

A gel-tube (a gel in silicone tubing) is pre-attachedto the C-Tube LCDM. Preparation is easy.

The crystals grown in the cell are stable in long term.
Using the gel soaking bag and the crystallization bag, you
can perform crystallization experiments in a small space.

How to assemble C-Tube LCDM
■Gel soaking

C-Tube LCDM should beimmersed into the proper gelsoaking solution in several
days before loading protein
solution. The gel-soaking bag
is sealed by a heat sealer(option).

■Solution preparation

Fill the crystallization bag with
0.5 mL of reservoir solutionusing a PTFE tubing tip.

■Crystallization cell set-up completedInsert the C-Tube LCDM
into the crystallization bag.

■Protein solution loading■Picking up C-Tube LCDM

After the gel-tube is wellsoaked with the gel-soaking
solution, pick up C-Tube
LCDM from the gel-soaking
solution.

■Sealing the bag

Load the protein solution into C-Tube LCDM
with the fine top tip, and plug the upper edge ofC-Tube LCDM into the C-Cap.

■Start crystallization

The crystallization bag
is sealed by the heatsealer.

■Instructional videoObserve crystal growth.

Related item ‘C-Chip-DM’ : Gel-tube with dialysis membrane■ MB2004-CRT901 C-Chip-DM15/ 6 pieces■ Features1.

Crystallization reagent gradually diffuses into a capillary through the gel-tube. Mild
crystallization condition can be realized.

2.

Small amount of sample is required in this dialysis method.
・0.3mm I.D. and 30mm length in a capillary: 2.2μl
・0.5mm I.D. and 40mm length in a capillary: 7.9μl

Patent No. JP 6473788
A part of this product utilizes the result of large crystal container improvement examination work contracted from JAXA.

Please read the instruction manual carefully before using this product.

Sales agent

Manufactured by

Confocal Science Inc.
Toyo 2-chome, Koto-ku, Tokyo
135-0016 JAPANhttp://www.cosmobio.co.jp/index_e.asp
E-mail: export@cosmobio.co.jp

Hayakawa 2nd Bldg 7F, 2-12-2 Iwamoto-cho,Chiyoda-ku, Tokyo 101-0032 JAPANhttp://www.confsci.co.jp/index_e.html
E-mail info@confsci.co.jp

In alliance with

Maruwa Foods and Biosciences Inc.
170-1 Tsutsui-cho, Yamatokoriyama-shi, Nara
639-1123 JAPANhttp://maruwafoods.jp/●The contents of the catalog are subject to change without notice●Printed in June 2019.