Navigation Links
Continuous-Elution Electrophoresis for Purification of the Baculovirus-Expressed Coronavirus Structural Proteins, Rev A

tting using porcine hyperimmune antiserum to TGEV (Stott, 1989) or a TGEV neutralizing monoclonal antibody specific for epitope A of the S-protein (25C9) and non-neutralizing monoclonal antibodies specific for epitope D of the S-protein (44C11) and N-protein (25H7) (Lanza et al., 1995). The protein concentration was estimated with spectrophotometer readings at 280 nm.


Results
By extrapolating the molecular size to prestained protein standards or to native coronavirus proteins from virus (Wesley et al., 1991) cell culture adapted TGEV (Miller strain), the recombinant TGEV M-protein (2831 kDa) was expected to be eluted within fraction range 913 (Figure 1A). Fractions 913 were pooled and concentrated. The presumed M-protein was electrophoresed and detected by Coomassie blue stain (Figure 1B, lane 5) and its virus specificity was confirmed by western blot (Figure 1C, lane 5). After concentration of fractions 913, the final protein concentration was 0.4 mg/ml or 5% of the initial solubilized cell lysate protein.

The recombinant TGEV N-protein (47 kDa) was expected to be within fraction range 2527 (Figure 2A). These fractions were pooled and concentrated. The concentrated sample was analyzed as described for the recombinant TGEV M-protein (Figure 2B, C). The resulting purified N-protein concentration was 0.5 mg/ml, or 8% of the initial baculovirus lysate.

The recombinant TGEV S-protein (150 kDa) was expected to be within fraction range 2030 (Figure 3A). Fractions 2030 were pooled and concentrated. The concentrated S-protein sample was further analyzed as described for the M- and N-proteins (Figure 3B, C). In addition to the major band of 150 kDa,
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. A Versatile Power Supply for All Electrophoresis Applications
2. A Multiple Mutation Model System as a Test Development and Training Tool for Denaturing Gradient Gel Electrophoresis
3. Detection of K-ras Point Mutations in the Pancreas by Constant Denaturing Gel Electrophoresis Using the DCode System
4. Detection of Mutations in the Fas Antigen of Lymphoma Tumors by RT-PCR and Denaturing Gradient Gel Electrophoresis
5. Detection of Variation in Highly Polymorphic Mhc Genes by Denaturing Gradient Gel Electrophoresis Using the DCode System
6. Detection of p53 Gene in Breast Cancer by Denaturing Gradient Gel Electrophoresis and the DCode System
7. Identification of Disease Causing Mutations in Phenylketonuria by Denaturing Gradient Gel Electrophoresis Using the DCode System
8. Microbial Diversity in Ground and Surface Water Analyzed by Denaturing Gradient Gel Electrophoresis Using the DCode System
9. Oligonucleotide Purity Analysis by Capillary Electrophoresis
10. Preparative Nondenaturing Gel Electrophoresis Used in the Purification of an Esterase Involved in Insecticide Resistance, Rev C
11. Preparative Nondenaturing Gel Electrophoresis to Purify NADP-Specific Glutamate Dehydrogenase From Chlorella, Rev B
Post Your Comments:
(Date:4/30/2015)... 2015 Nitto Avecia, a leading ... increased capacity and production capability to serve biotechnology ... and service fulfillment continues to drive Nitto Avecia ... throughput capacities. , In Fall 2014, ... process equipment at its Milford, MA facility, which ...
(Date:4/30/2015)... (PRWEB) April 30, 2015 Biodata, ... notebook with integrated protocols and specimen management systems, ... solutions and consulting services for life science research, ... of Labguru’s newest feature: chemical structure and reaction ... features for biologists, bio-chemists, and more, the integration ...
(Date:4/30/2015)... , April 30, 2015  Asterias Biotherapeutics, Inc. ... emerging field of regenerative medicine, today announced that it ... Thursday, May 7, 2015 after the close of the ... call and webcast on Thursday, May 7, 2015 at ... the results and recent corporate developments. For ...
(Date:4/30/2015)... 2015 Available in select spas ... , Fleur’s introduces CC CRÈME ... benefits of makeup, skin care and solar protection ... is designed with specific complexion-correcting pigments to instantly ... and a healthy glow. Far more than a ...
Breaking Biology Technology:Biotechnology Manufacturer Nitto Avecia Expands Capacity to Meet Client Demand 2Labguru Integrates ChemAxon's Marvin JS, to Enhance Functionality for Chemists 2Asterias Biotherapeutics to Report First Quarter Results on May 7, 2015 2Asterias Biotherapeutics to Report First Quarter Results on May 7, 2015 3Fleur’s Introduces CC CRÈME Perfect Skin Solution 2Fleur’s Introduces CC CRÈME Perfect Skin Solution 3
... accuracy: DNA vaccines that home in on DCs are ... develop new vaccines against infectious diseases is DNA vaccination. ... vaccine, the body converts the information in the DNA ... However, current DNA vaccines induce relatively weak immune responses ...
... UK, 6 March 2008 Elsevier, the world-leading ... and services, announced today Hybrid Materials 2009: The ... which will take place in Tours, France, 15-19 ... has experienced a 14% annual growth in published ...
... DNDN ) will release its fourth quarter financial results on,Thursday, March ... PT; 11:00 a.m. ET to review fiscal year 2007 financial results,and ... follows:, Time: 11:00 AM ... March 13, 2008 Dial-in: 1-877-548-7901 (domestic) or +1-719-325-4844 ...
Cached Biology Technology:JCI online early table of contents: March 6, 2008 2JCI online early table of contents: March 6, 2008 3JCI online early table of contents: March 6, 2008 4Hybrid materials: Exciting interdisciplinary field offering future solutions for industry 2-Dendreon to Hold Conference Call on Thursday, March 13, 2008 at 11:00 AM ET- 2
(Date:4/8/2015)... N.C. , April 8, 2015  Infinisource, ... solutions and services, and Morpho, a leading supplier ... partnership to produce the market,s most advanced biometrically-enabled ... Infinisource,s iSolved NXG time clock is setting a ... and labor data for the small and mid-size ...
(Date:4/2/2015)... , April 2, 2015 ... DERMALOG by far outperforms the strongest competitors, showing ... high accuracy, the system is capable of a ... speed proves up to be ten times faster ... its runner-up performance.      (Photo: ...
(Date:4/2/2015)... 2015 According to preliminary estimates ... somewhat higher than 125 MSEK. This exceeds the guidance ... that revenues for Q1 2015 would be in line ... Q4 2014. The operating result for the first quarter ... interim report will as previously communicated be published on ...
Breaking Biology News(10 mins):iSolved Time Clock Offers Unmatched Accuracy due to Morpho Biometrics 2iSolved Time Clock Offers Unmatched Accuracy due to Morpho Biometrics 3DERMALOG AFIS Emerges as Performance Winner in NIST Benchmark Test 2DERMALOG AFIS Emerges as Performance Winner in NIST Benchmark Test 3Fingerprint Cards Revises Revenues for Q1 2015 2
... Des Moines, Iowa, USA March 18, 2008 A ... of Rwandan rain forest will have a greater range ... of Africas most ambitious forest restoration and ecological research efforts ... Park, said today that a 30-mile (50km) tree corridor will ...
... the United Kingdom and France have identified four ... of Creutzfeldt-Jakob disease. The study, published March 14th ... these subgroups could represent distinct prion strains in ... Prion diseases are transmissible neurodegenerative disorders characterized ...
... Take a deers body, attach a camels head and add ... odd-ball antelope with the enormous schnoz that lives on the isolated ... they are strange-looking due to over-hunting. Now, according to a ... as well. ...
Cached Biology News:Rwanda conservation effort to link isolated chimps to distant forest 2Rwanda conservation effort to link isolated chimps to distant forest 3Asia's odd-ball antelope faces migration crisis 2
... The ULS cDNA Synthesis and Labeling Kit ... hybridization experiments. First unmodified cDNA is made ... cDNA using an Oligo(dT) primer. Unlike other ... used in the reverse transcription reaction. This ...
... Dot Blot Kits overcome this limitation and ... a wide range in low-density protein array ... They are ideal for performing reverse-phase protein ... for the presence of low abundant antigens. ...
AC input: 50-60 Hz230 V...
... Convenient, easy-to-use kits for arraying ... detection using common lab equipment. DiscoverLight ... analysis of your protein, cell lysate ... format. The kits include specially designed ...
Biology Products: