Rapid and reliable preparation of total cell lysates from fission yeast

Author(s):  
Atsuko Shirai ◽  
Akihisa Matsuyama ◽  
Yoko Yashiroda ◽  
Ritsuko Arai ◽  
Minoru Yoshida
2014 ◽  
Vol 10 (1) ◽  
pp. 134-148 ◽  
Author(s):  
Maya Tanase ◽  
Valerio Zolla ◽  
Cristina C Clement ◽  
Francesco Borghi ◽  
Aleksandra M Urbanska ◽  
...  

2019 ◽  
Author(s):  
Zachary VanAernum ◽  
Florian Busch ◽  
Benjamin J. Jones ◽  
Mengxuan Jia ◽  
Zibo Chen ◽  
...  

It is important to assess the identity and purity of proteins and protein complexes during and after protein purification to ensure that samples are of sufficient quality for further biochemical and structural characterization, as well as for use in consumer products, chemical processes, and therapeutics. Native mass spectrometry (nMS) has become an important tool in protein analysis due to its ability to retain non-covalent interactions during measurements, making it possible to obtain protein structural information with high sensitivity and at high speed. Interferences from the presence of non-volatiles are typically alleviated by offline buffer exchange, which is timeconsuming and difficult to automate. We provide a protocol for rapid online buffer exchange (OBE) nMS to directly screen structural features of pre-purified proteins, protein complexes, or clarified cell lysates. Information obtained by OBE nMS can be used for fast (<5 min) quality control and can further guide protein expression and purification optimization.


2013 ◽  
Vol 5 (1) ◽  
Author(s):  
Lies Indah Sutiknowati

The objective of this research was to evaluate waters quality in Pari island waters for aquaculture purpose based on bacteriological information conducted in Mei and September 2011. Microbiological parameters analyzed were total density of bacteria for coliforms, E.coli, pathogenic, heterotrophic, halotoleran, phosphate-nitrate-ammonia breaker, and total cells. Method to analyze coliform bacteria was filtration, identification of pathogenic bacteria using biochemical test, density analises for heterotrophic bacteria, analises for phosphate-nitrate-ammonia breaker bacteria using pour plate, and total cell using Acridine Orange Epifluorescence Microscopy. Results showed that the abundance of total coliform cell was about 1000-7000 colony forming unit (cfu)/100 ml. The abundance of heterotrophic, halotolerant, phosphate-nitrate-ammonia bacteria in seawater was (3.6-4.3)x105 cfu/ml, (1.1-1.3)x105 cfu/ml, (0.5-3.44)x103cfu/ml; and (3.6-6.7)x105 cfu/ml, (1.6-2.7)x105 cfu/ml, (0.6-5.22)x103 cfu/ml in sediment, respectively. The total cell of bacteria was (0.05-2.1)x107cells/ml. The dog-conch (Strombus turturella) and blood-clamps (Anadara granosa) can survive in Pari Island and there was a significant increase in sea grass litter with growth average of 0.67 mm/day and 0.90 mm/day. During snails and clamps growth, there were found several genus of pathogenic bacteria such as Salmonella, Vibrio, Aeromonas, Pseudomonas, Citrobacter, Proteus, Shigella, Hafnia, and Yersinia. The results showed that Pari island waters was suitable for developing shellfish aquaculture dog conch and blood clamps. Keywords: bacteria, parameter, shellfish, aquaculture.


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