Growth of large protein crystals by a large-scale hanging-drop method

2010 ◽  
Vol 43 (4) ◽  
pp. 937-939 ◽  
Author(s):  
Keisuke Kakinouchi ◽  
Tsutomu Nakamura ◽  
Taro Tamada ◽  
Hiroaki Adachi ◽  
Shigeru Sugiyama ◽  
...  
Keyword(s):  
Large Scale ◽  
Protein Crystals ◽  
Hanging Drop ◽  
Large Protein ◽  
Drop Method ◽  
Egg White Lysozyme ◽  
Crystallization Experiments ◽  
Pipette Tip ◽  
Hen Egg White ◽  
Hanging Drop Method

A method for growing large protein crystals is described. In this method, a cut pipette tip is used to hang large-scale droplets (maximum volume 200 µl) consisting of protein and precipitating agents. A crystal grows at the vapor–liquid interface; thereafter the grown crystal can be retrieved by droplet–droplet contact both for repeated macroseeding and for mounting crystals in a capillary. Crystallization experiments with peroxiredoxin ofAeropyrum pernixK1 (thioredoxin peroxidase, ApTPx) and hen egg white lysozyme demonstrated that this large-scale hanging-drop method could produce a large-volume crystal very effectively. A neutron diffraction experiment confirmed that an ApTPx crystal (6.2 mm3) obtained by this method diffracted to beyond 3.5 Å resolution.

A simple technique to convert sitting-drop vapor diffusion into hanging-drop vapor diffusion by solidifying the reservoir solution with agarose

2009 ◽  
Vol 42 (5) ◽  
pp. 975-976 ◽  
Author(s):  
Tae Woong Whon ◽  
Yong-Hwan Lee ◽  
Dong-Shan An ◽  
Hyun Kyu Song ◽  
Song-Gun Kim
Keyword(s):  
Protein Crystallization ◽  
Glucose Isomerase ◽  
Diffusion Experiment ◽  
Hanging Drop ◽  
Vapor Diffusion ◽  
Egg White Lysozyme ◽  
Simple Protocol ◽  
Crystallization Experiments ◽  
Reservoir Solution ◽  
Hen Egg White

A simple protocol to convert sitting-drop vapor-diffusion plating into a hanging-drop vapor-diffusion experiment in protein crystallization is reported. After making a sitting-drop plate, agarose solution was added to solidify the reservoir solution, and the plates were incubated upside down. Crystallization experiments with hen egg white lysozyme, thaumatin and glucose isomerase showed that the `upside-down sitting-drop' method could produce single crystals with all the benefits of the hanging-drop crystallization method.

scholarly journals Diffraction study of protein crystals grown in cryoloops and micromounts

2010 ◽  
Vol 43 (6) ◽  
pp. 1513-1518 ◽  
Author(s):  
Michael A. Berger ◽  
Johannes H. Decker ◽  
Irimpan I. Mathews
Keyword(s):  
Data Collection ◽  
Diffraction Study ◽  
New Method ◽  
Protein Crystals ◽  
Hanging Drop ◽  
Drop Method ◽  
Laboratory Conditions ◽  
Crystallization Conditions ◽  
Diffraction Patterns ◽  
Hanging Drop Method

Protein crystals are usually grown in hanging or sitting drops and generally get transferred to a loop or micromount for cryocooling and data collection. This paper describes a method for growing crystals on cryoloops for easier manipulation of the crystals for data collection. This study also investigates the steps for the automation of this process and describes the design of a new tray for the method. The diffraction patterns and the structures of three proteins grown by both the new method and the conventional hanging-drop method are compared. The new setup is optimized for the automation of the crystal mounting process. Researchers could prepare nanolitre drops under ordinary laboratory conditions by growing the crystals directly in loops or micromounts. As has been pointed out before, higher levels of supersaturation can be obtained in very small volumes, and the new method may help in the exploration of additional crystallization conditions.

Fabrication of three-dimensional islet models by the geometry-controlled hanging-drop method

2019 ◽  
Vol 35 (2) ◽  
pp. 329-337 ◽  
Author(s):  
Bin Gao ◽  
Ce Jing ◽  
Kelvin Ng ◽  
Belinda Pingguan-Murphy ◽  
Qingzhen Yang
Keyword(s):  
Three Dimensional ◽  
Hanging Drop ◽  
Drop Method ◽  
Hanging Drop Method

Retracted: Mouse ES cells have a potential to differentiate into odontoblast‐like cells using hanging drop method

Oral Diseases ◽  
2013 ◽  
Vol 20 (4) ◽  
pp. 395-403 ◽  
Author(s):  
R Kawai ◽  
N Ozeki ◽  
H Yamaguchi ◽  
T Tanaka ◽  
K Nakata ◽  
...  
Keyword(s):  
Es Cells ◽  
Hanging Drop ◽  
Drop Method ◽  
Mouse Es Cells ◽  
Hanging Drop Method

Enrichment of cardiac differentiation of mouse embryonic stem cells by optimizing the hanging drop method

2010 ◽  
Vol 33 (4) ◽  
pp. 853-858 ◽  
Author(s):  
Ming Chen ◽  
Yong-Qing Lin ◽  
Shuang-Lun Xie ◽  
Hong-Fu Wu ◽  
Jing-Feng Wang
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells ◽  
Cardiac Differentiation ◽  
Hanging Drop ◽  
Drop Method ◽  
Hanging Drop Method

Digital topography with an X-ray CCD camera for characterizing perfection in protein crystals

2012 ◽  
Vol 45 (5) ◽  
pp. 1009-1014 ◽  
Author(s):  
Kei Wako ◽  
Kunio Kimura ◽  
Yu Yamamoto ◽  
Takuya Sawaura ◽  
Mengyuan Shen ◽  
...  
Keyword(s):  
Ccd Camera ◽  
Egg White ◽  
Protein Crystals ◽  
Hen Egg White Lysozyme ◽  
X Ray ◽  
Crystal Perfection ◽  
Egg White Lysozyme ◽  
Domain Structures ◽  
Peak Broadening ◽  
Hen Egg White

Digital X-ray topography using an X-ray CCD camera and conventional X-ray topography using X-ray film were used to investigate tetragonal hen egg-white lysozyme (HEWL) crystals. Previously, clear dislocation images of protein crystals were mainly obtained by film methods. Earlier studies of HEWL crystals using an X-ray CCD camera mainly revealed domain structures. In the present study, dislocation images of the same HEWL crystal have been obtained by using conventional X-ray film and a digital X-ray CCD camera. The results demonstrate that digital topography using an X-ray CCD camera is an effective method for characterizing protein crystals. A series of digital topographic images were analyzed by the method developed by Lovelace, Murphy, Pahl, Brister & Borgstahl [J. Appl. Cryst.(2006),39, 425–432]. Sub-peaks and peak broadening originating from dislocations in local rocking curves were observed. Moreover, the crystal perfection was evaluated by mapping the angular positions of the maximums and the full widths at half-maximum of local rocking curves.

Development of Artificial Blood Clot by Hanging Drop Method for Retinal Vascular Occlusion

2018 ◽  
Vol 2018.9 (0) ◽  
pp. 30pm4PN38
Author(s):  
Daisuke MATSUMURA ◽  
Mei SUMIMOTO ◽  
Keiko MIWA ◽  
Hideyasu OH ◽  
Yasuhiro MORIIZUMI ◽  
...  
Keyword(s):  
Blood Clot ◽  
Vascular Occlusion ◽  
Hanging Drop ◽  
Artificial Blood ◽  
Drop Method ◽  
Retinal Vascular Occlusion ◽  
Hanging Drop Method
Sign in / Sign up

Export Citation Format

Share Document