Blotting from Immobilized pH Gradient Gels: Application to Total Cell Lysates

Author(s):  
Harry Towbin
Keyword(s):  
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.


2019 ◽  
Vol 16 (10) ◽  
pp. 940-950 ◽  
Author(s):  
Jiandong Yu ◽  
Zhi Chen ◽  
Yan-zhi Yin ◽  
Chaoyuan Tang ◽  
Enying Hu ◽  
...  

Background: In this study, a liposomal gel based on a pH-gradient method was used to increase the skin-layer retention of monocrotaline (MCT) for topical administration. Methods: Using the Box-Behnken design, different formulations were designed to form liposome suspensions with optimal encapsulation efficiency (EE%) and stability factor (KE). In order to keep MCT in liposomes and accumulate in skin slowly and selectively, MCT liposome suspensions were engineered into gels. Results: A pH-gradient method was used to prepare liposome suspensions. The optimal formulation of liposome suspensions (encapsulation efficiency: 83.10 ± 0.21%) was as follows: MCT 12 mg, soybean phosphatidyl choline (sbPC) 200 mg, cholesterol (CH) 41 mg, vitamin E (VE) 5 mg, and citric acid buffer solution (CBS) 4.0 10 mL (pH 7.0). The final formulation of liposomal gels consisted of 32 mL liposome suspensions, 4.76 mL deionized water, 0.40 g Carbopol-940, 1.6 g glycerol, 0.04 g methylparaben, and a suitable amount of triethanolamine for pH value adjustment. The results of in vitro drug release showed that MCT in liposomal gels could be released in 12 h constantly in physiological saline as a Ritger-Peppas model. Compared with plain MCT in gel form, liposomal MCT in gel had higher skin retention in vitro. Conclusion: In this study, liposomal gels were formed for greater skin retention of MCT. It is potentially beneficial for reducing toxicities of MCT by topical administration with liposomal gel.


Sign in / Sign up

Export Citation Format

Share Document