The two-dimensional gel electrophoresis (2D-GE) is a technique that is very necessary for the study of biological system proteomas, providing an assessment of changes in abundance of proteins in various experimental conditions. However, because of the complexity of 2D-GE gel, there is no systematic, automatic, and reproducing protocol for image analysis and specific implementation is needed for each context. In addition, practically all available solutions are commercial, which implies high costs and a little flexibility to modulate algorithm parameters.
Using bacterial strains, pseudomonas aeruginosaag1 as a model, we get pictures of 2D-GE protein protein profiles periplasm when strains are exposed to several conditions, including antibiotics. Then, we proceed to implement and evaluate the image analysis protocol with open source software, CellProfiler. First, step preprocessing includes bunwarpj-image pipes to align 2D-GE images. Then, using CellProfiler, we standed two pipes for place identification. The total introduction of places is achieved by using segmentation with intensity, which performance is evaluated when compared to the reference protocol.
In the second pipe with the same program, the identification of differential spots is handled when comparing pairs of protein profiles. Because the program characteristics used, our workflow can automatically analyze a large number of images and parallel, which is an advantage in connection with other implementations. Finally, we compare six experimental conditions of bacterial pressure in the presence or absence of antibiotics, determine the relationship of protein profiles by applying the PCA clustering algorithm (main component analysis) and hierarchy (clustering).
Electrophoresis gel polyacrylamide
Chains of cross-linked from polyacrylamides can be used as electric neutral gel to separate multiple-double DNA fragments according to the size and DNAS stranded single according to size and conformation. Polyacrylamide gel has three main advantages along with axible glass: (1) the power of resolution is so large that they can separate the DNA molecules that are different in length to only 0.1% (eg) 1000 bp). (2) They can accommodate the amount of DNA which is far greater than agarose gel. Up to 10 μg of DNA can be applied to one slot (1 cm × 1 mm) from a typical polyacrylamide gel without losing resolution. (3) DNA recovers from polyacrylamide gel is very pure and can be used for the most demanding purpose (eg, microinjection embryo mouse).
However, polyacrylamide gel has losses to be more difficult to be prepared and handled than gel agarose. The method is presented here to prepare and run polyacrylamide gels that are not prominent and for DNA detection on this gel with coloring.
Comparative analysis using electrophoresis of pulsating gel-field highlights the transmission of salmonella potential between buffalo and asimptomatic pigs on one agriculture
Buffalo and pigs play an important epidemiological roll in the cycle of salmonella infection, and asymptomatic animals can act as a key component in the spread of this disease with horizontal, vertical, and cross species transmissions. Our research is intended and able to confirm the evidence of transmission of cross-agona salmonella species between buffalo and asimptomatic pigs. Also, we describe salmonella infection in the pork production phase, which involves Serotype Agona, Senftenberg and Schwarzengrund.
Rectal samples are collected from buffalo jafarabadi (n = 25) and pig piau (n = 32), located on an agriculture. Salmonella Agona was isolated from the buffalo of breastfeeding, gilt, sowing pregnant, and weaning pigs, Salmonella Schwarzengrund from sowing breastfeeding and Salmonella Senftenberg from Gilt, sowing pregnant, sowing weaning, and weaning pigs. The pulsed gel electrophoresis protocol (PFGE) is done and reveals four different profiles. Profile 1 (Salmonella Agona), isolated from pregnant pigs, gold and two breastfeeding buffalo, revealing the PFGE pattern that cannot be distinguished, confirming proof of cross-species transmission potential. Profile 2 (Salmonella Agona), 3 (Salmonella Senftenberg), and 4 (Salmonella Schwarzengrund), isolated from pigs, revealing pfge patterns that cannot be distinguished, proving salmonella infection in the pork production phase.
Consider the epidemiological relevance of the buffalo and pigs in the cycle of salmonella infection, transmission confirmation of the potential species of salmonella agona and potential salmonella infections in the pork production phase highlight the importance of the formation of preventive health strategies correctly on animal husbandry, in particular the importance of avoiding contact between buffalo and pigs, due to cross transmission Species can occur, increasing the risk of disseminating disease.
Seabeam wild or farming gilead (Sparus Aurata)? How to distinguish between them with two-dimensional gel electrophoresis
Because the shares of world wild fish are limited and market demand increases, fish farming has become an alternative food source and a way to reduce costs for consumers. Agricultural sales as wild fish are fraud practices, because it is important to work to provide new tools and alternatives that can help in combating fraud to protect consumers and to ensure food tracing. Proteomic profiles of farm fish and wildly different from each other. With this study we want to identify the marker of the liver protein through two-dimensional electrophoresis (2DE) which will enable us to distinguish wildly from the Seabream Gilhead farming.
 ENDURO 15.10 Horizontal Electrophoresis 15 x 10cm Gels |
|
LE1004 |
GenDepot |
Set |
EUR 886.8 |
 ENDURO 15.15 Horizontal Electrophoresis 15 x 15cm Gels |
|
LE1005 |
GenDepot |
Set |
EUR 1081.2 |
 ENDURO 20.20 Horizontal Electrophoresis 20 x 20cm Gels |
|
LE1006 |
GenDepot |
Set |
EUR 1170 |
 runVIEW horizontal electrophoresis system 15 x 7cm gel tray 1 set of casting dams 8 double sided combs - EACH |
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ELE2300 |
Scientific Laboratory Supplies |
EACH |
EUR 1802.25 |
 runVIEW horizontal electrophoresis system with 15 x 10cm gel tray 1 set of casting dams and 8 double sided combs - EACH |
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ELE2302 |
Scientific Laboratory Supplies |
EACH |
EUR 1891.35 |
 Glycine for electrophoresis, 99.5% |
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52574 |
Sisco Laboratories |
100 Gms |
EUR 9.58 |
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Description: Part A |
 Agarose for DNA electrophoresis |
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16-0027-500 |
Medicago |
500 g |
EUR 410.4 |
 Aminoacetic acid, ?For electrophoresis |
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GRM2233-100G |
EWC Diagnostics |
1 unit |
EUR 32.26 |
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Description: Aminoacetic acid, ?For electrophoresis |
Aminoacetic acid, ?For electrophoresis |
|
GRM2233-500G |
EWC Diagnostics |
1 unit |
EUR 144.96 |
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Description: Aminoacetic acid, ?For electrophoresis |
 Agarose, medium EEO ?For electrophoresis |
|
RM6249-100G |
EWC Diagnostics |
1 unit |
EUR 119.06 |
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Description: Agarose, medium EEO ?For electrophoresis |
Agarose, medium EEO ?For electrophoresis |
|
RM6249-10G |
EWC Diagnostics |
1 unit |
EUR 19.63 |
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Description: Agarose, medium EEO ?For electrophoresis |
Agarose, medium EEO ?For electrophoresis |
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RM6249-1KG |
EWC Diagnostics |
1 unit |
EUR 1089.43 |
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Description: Agarose, medium EEO ?For electrophoresis |
Agarose, medium EEO ?For electrophoresis |
|
RM6249-25G |
EWC Diagnostics |
1 unit |
EUR 36.87 |
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Description: Agarose, medium EEO ?For electrophoresis |
 Electrophoresis Module for Eco-Mini, for 2 gels |
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AJ846-017-175 |
Westburg |
each |
EUR 604.95 |
 Electrophoresis Module for Eco-Maxi, for 2 gels |
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AJ846-017-475 |
Westburg |
each |
EUR 997.35 |
 ES 6 Tooth Gel Comb For ES Electrophoresis |
|
LE1011-A4 |
GenDepot |
Ea |
EUR 148.8 |
 for electrophoresis, 99%) Ammonium Persulphate (APS) for electrophoresis, 99% |
|
28575 |
Sisco Laboratories |
25 Gms |
EUR 1.51 |
|
|
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Description: Part A |
 ES 8/10 Double Gel Comb For ES Electrophoresis |
|
LE1011-A3 |
GenDepot |
Ea |
EUR 148.8 |
 Tris-HEPES-SDS Buffer 20X for Electrophoresis |
|
20840057-1 |
Glycomatrix |
500 mL |
EUR 74.66 |
Tris-HEPES-SDS Buffer 20X for Electrophoresis |
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20840057-2 |
Glycomatrix |
1 L |
EUR 139.51 |
 electrophoresis ps 1500v, 300ma, 150w |
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EV2230 |
Consort |
ea |
EUR 898.8 |
electrophoresis ps 1500v, 300ma, 150w |
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EV2230-UK |
Consort |
ea |
EUR 898.8 |
electrophoresis ps 1500v, 300ma, 150w |
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EV2230-US |
Consort |
ea |
EUR 898.8 |
 electrophoresis ps 3000v, 150ma, 150w |
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EV2320 |
Consort |
ea |
EUR 898.8 |
electrophoresis ps 3000v, 150ma, 150w |
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EV2320-UK |
Consort |
ea |
EUR 898.8 |
electrophoresis ps 3000v, 150ma, 150w |
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EV2320-US |
Consort |
ea |
EUR 898.8 |
 electrophoresis ps 1200v, 500ma, 300w |
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EV3150 |
Consort |
ea |
EUR 960 |
electrophoresis ps 1200v, 500ma, 300w |
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EV3150-UK |
Consort |
ea |
EUR 960 |
electrophoresis ps 1200v, 500ma, 300w |
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EV3150-US |
Consort |
ea |
EUR 960 |
 electrophoresis ps 3000v, 300ma, 300w |
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EV3330 |
Consort |
ea |
EUR 1456.8 |
electrophoresis ps 3000v, 300ma, 300w |
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EV3330-UK |
Consort |
ea |
EUR 1456.8 |
electrophoresis ps 3000v, 300ma, 300w |
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EV3330-US |
Consort |
ea |
EUR 1456.8 |
 electrophoresis ps 6000v, 150ma, 300w |
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EV3620 |
Consort |
ea |
EUR 1456.8 |
electrophoresis ps 6000v, 150ma, 300w |
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EV3620-UK |
Consort |
ea |
EUR 1456.8 |
electrophoresis ps 6000v, 150ma, 300w |
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EV3620-US |
Consort |
ea |
EUR 1456.8 |
) Electrophoresis Buffer (10x) |
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MBS2557002-100mL |
MyBiosource |
100mL |
EUR 85 |
Electrophoresis Buffer (10x) |
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MBS2557002-200mL |
MyBiosource |
200mL |
EUR 90 |
Electrophoresis Buffer (10x) |
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MBS2557002-500mL |
MyBiosource |
500mL |
EUR 95 |
Electrophoresis Buffer (10x) |
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MBS2557002-5x500mL |
MyBiosource |
5x500mL |
EUR 380 |
 electrophoresis ps 400v, 500ma, 50w |
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EV1450 |
Consort |
ea |
EUR 574.8 |
electrophoresis ps 400v, 500ma, 50w |
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EV1450-UK |
Consort |
ea |
EUR 574.8 |
electrophoresis ps 400v, 500ma, 50w |
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EV1450-US |
Consort |
ea |
EUR 574.8 |
 electrophoresis ps 300v, 1000ma, 150w |
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EV2310 |
Consort |
ea |
EUR 686.4 |
electrophoresis ps 300v, 1000ma, 150w |
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EV2310-UK |
Consort |
ea |
EUR 686.4 |
electrophoresis ps 300v, 1000ma, 150w |
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EV2310-US |
Consort |
ea |
EUR 686.4 |
 electrophoresis ps 600v, 500ma, 150w |
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EV2650 |
Consort |
ea |
EUR 686.4 |
electrophoresis ps 600v, 500ma, 150w |
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EV2650-UK |
Consort |
ea |
EUR 686.4 |
electrophoresis ps 600v, 500ma, 150w |
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EV2650-US |
Consort |
ea |
EUR 686.4 |
 electrophoresis ps 300v, 2000ma, 300w |
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EV3020 |
Consort |
ea |
EUR 960 |
electrophoresis ps 300v, 2000ma, 300w |
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EV3020-UK |
Consort |
ea |
EUR 960 |
electrophoresis ps 300v, 2000ma, 300w |
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EV3020-US |
Consort |
ea |
EUR 960 |
 electrophoresis ps 600v, 1000ma, 300w |
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EV3610 |
Consort |
ea |
EUR 960 |
electrophoresis ps 600v, 1000ma, 300w |
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EV3610-UK |
Consort |
ea |
EUR 960 |
electrophoresis ps 600v, 1000ma, 300w |
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EV3610-US |
Consort |
ea |
EUR 960 |
 TEMED, electrophoresis grade |
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RM10358-100G |
EWC Diagnostics |
1 unit |
EUR 301.18 |
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Description: TEMED, electrophoresis grade |
 Protein Antioxidant for SDS-PAGE Gel Electrophoresis |
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21440026-1 |
Glycomatrix |
15 mL |
EUR 17.3 |
Protein Antioxidant for SDS-PAGE Gel Electrophoresis |
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21440026-2 |
Glycomatrix |
25 mL |
EUR 23.23 |
Protein Antioxidant for SDS-PAGE Gel Electrophoresis |
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21440026-3 |
Glycomatrix |
50 mL |
EUR 35.42 |
Protein Antioxidant for SDS-PAGE Gel Electrophoresis |
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21440026-4 |
Glycomatrix |
100 mL |
EUR 62.99 |
 Electrophoresis Power Supply |
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LA690-1NO |
EWC Diagnostics |
1 unit |
EUR 432.84 |
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Description: Electrophoresis Power Supply |
 electrophoresis power supply |
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EV245 |
Consort |
ea |
EUR 574.8 |
 Agarose Gel Electrophoresis Kit |
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MB570-10PR |
EWC Diagnostics |
1 unit |
EUR 38.47 |
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Description: Agarose Gel Electrophoresis Kit |
 10X MOPS Electrophoresis Buffer |
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ML050-500ML |
EWC Diagnostics |
1 unit |
EUR 32.7 |
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Description: 10X MOPS Electrophoresis Buffer |
 CometAssay Electrophoresis System |
|
4250-050-ES |
Biotechne |
1 Kit |
EUR 3540.96 |
 myGel Mini Electrophoresis System |
|
BM0286 |
GenDepot |
System |
EUR 741.6 |
 SDS-PAGE Electrophoresis Buffer |
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MBS355536-1L |
MyBiosource |
1L |
EUR 165 |
SDS-PAGE Electrophoresis Buffer |
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MBS355536-5x1L |
MyBiosource |
5x1L |
EUR 480 |
SDS-PAGE Electrophoresis Buffer |
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MBS355542-10x1L |
MyBiosource |
10x1L |
EUR 250 |
 6X6 well Electrophoresis Apparatus |
|
EDK626 |
Bio Basic |
1 UNIT |
EUR 750.6 |
|
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 electrophoresis power supply 120 vac |
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EV245-US |
Consort |
ea |
EUR 574.8 |
 electrophoresis power supply uk plug |
|
EV245-UK |
Consort |
ea |
EUR 574.8 |
 CometAssay Electrophoresis Starter Kit |
|
4250-050-ESK |
Biotechne |
1 Kit |
EUR 3749.28 |
 Alkaline Hemoglobin Electrophoresis Kit |
|
MBP001-112RUNS |
EWC Diagnostics |
1 unit |
EUR 118.87 |
|
Description: Alkaline Hemoglobin Electrophoresis Kit |
Alkaline Hemoglobin Electrophoresis Kit |
|
MBP001-224RUNS |
EWC Diagnostics |
1 unit |
EUR 214 |
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Description: Alkaline Hemoglobin Electrophoresis Kit |
Alkaline Hemoglobin Electrophoresis Kit |
|
MBP008-224RUNS |
EWC Diagnostics |
1 unit |
EUR 208.42 |
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Description: Alkaline Hemoglobin Electrophoresis Kit |
 Alkaline Paper Electrophoresis Kit (Cell |
|
MBP009-20RUNS |
EWC Diagnostics |
1 unit |
EUR 59.3 |
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Description: Alkaline Paper Electrophoresis Kit (Cell |
 WEE VERT PROTEIN ELECTROPHORESIS SYSTEM |
|
LA1070-1NO |
EWC Diagnostics |
1 unit |
EUR 1258.71 |
|
Description: WEE VERT PROTEIN ELECTROPHORESIS SYSTEM |
 Electrophoresis Power Supply (2 termina |
|
LA842-1NO |
EWC Diagnostics |
1 unit |
EUR 66.05 |
|
Description: Electrophoresis Power Supply (2 termina |
 Cellulose Acetate Electrophoresis System |
|
LA1027-1NO |
EWC Diagnostics |
1 unit |
EUR 534.37 |
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Description: Cellulose Acetate Electrophoresis System |
) Agarose Gel Electrophoresis Kit (Teaching) |
|
66602 |
Sisco Laboratories |
15 expt. Kit |
EUR 44.05 |
|
|
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Description: Part E |
 ES Electrophoresis Casting Tray, 7cm X 7cm |
|
LE1011-A2 |
GenDepot |
Ea |
EUR 181.2 |
 20X DoubleQuick DNA Electrophoresis Buffer |
|
73530 |
Sisco Laboratories |
250 ml |
EUR 35.58 |
|
|
|
Description: Part E |
20X DoubleQuick DNA Electrophoresis Buffer |
|
73530-1 |
Sisco Laboratories |
500 ml |
EUR 64.4 |
|
|
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Description: Part E |
 HiPer® Agarose Gel Electrophoresis Teach |
|
HTBM001-10PR |
EWC Diagnostics |
1 unit |
EUR 29.56 |
|
Description: HiPer® Agarose Gel Electrophoresis Teach |
 ES Electrophoresis Casting Tray, 7cm X 14cm |
|
LE1011-A1 |
GenDepot |
Ea |
EUR 198 |
) Electrophoresis Apparatus - 7x10cm (6-10-well) |
|
EDK611 |
Bio Basic |
1 UNIT |
EUR 383.11 |
|
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) Electrophoresis Apparatus - 7x14cm (6-10-well) |
|
EDK613 |
Bio Basic |
1 UNIT |
EUR 487.51 |
|
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 CometAssay Single Cell Gel Electrophoresis Assay |
|
4250-050-K |
Biotechne |
50 tests |
EUR 513.6 |
Heart samples of 32 Gilead Seabream, wild and farming, stored at -80 ° C Previous extraction of protein. Samples are subject to 2DE to detect qualitative and quantitative differences. Proteom analysis shows a protein point (molecular weight ~ 34 kDa and isoelectric point ~ 6.9) only in the sample of the Seabread Wild Gilead; Liquid chromatography Tandem mass spectrometry identifies the place as ubiquitin. Ubiquitin can be a valid marker to distinguish wildly from the Seabread of Gilead farming, thus ensuring sustainable monitoring throughout the commercial chain and fighting commercial fraud.
