The molecular mechanisms whereby placental progress issue (PlGF) mediates its results in nonproliferative diabetic retinopathy (DR) are unknown. To raised perceive the position of PlGF in DR, we used tandem mass tags (TMT)-labeled quantitative proteomics to human retinal endothelial cells (HRECs), handled anti-PlGF antibody, and PBS as a management. Practical annotation and pathway enrichments have been carried out, which instructed that the differentially expressed proteins (DEPs) have been concerned in key metabolic processes, protein binding, and membrane, pentose phosphate pathway PPP and adherens junction. We performed built-in gene profiles of our beforehand printed transcriptomic knowledge to the TMT-labeled proteomics knowledge.
The outcomes confirmed the sixty proteins have been discovered to be modified on the mRNA ranges. The purposeful annotation performed for the sixty proteins instructed that 58.3% of proteins have been concerned in PPP, 25% of proteins have been in interleukin-12 singling and 16.7% of proteins have been concerned in glycolysis and gluconeogenesis pathway. Mass spec outcomes have been validated by transendothelial electrical resistance measurement by {an electrical} cell-impedance sensing and western blot evaluation of VE-cadherin, G6PD. These findings counsel that the PPP proteins and antioxidants might act as a downstream goal of PlGF and should play a decisive position in HREC organic features in DR.
PlGF (Placental progress issue) is understood to play a pivotal position in pathological angiogenesis and irritation by stimulating endothelial cell migration and by recruiting pericytes and inflammatory cells comparable to microglia and macrophages. Regardless of the well-defined pathophysiological roles of PlGF, the underlying molecular and mobile mechanisms usually are not utterly understood, particularly the precise relationships between biochemical occasions and molecular pathways regulated by PlGF, whose inhibition displays a protecting position in diabetic retinopathy.
This examine supplies new insights into protein expression patterns and permits the identification of many enticing candidates for investigation of PPP pathway position within the activation of the antioxidant protection system in diabetic retinopathy (DR). Our findings counsel that the PPP proteins and antioxidants (PRDX6, HMOX1, NQO1 and YES1) might act as downstream targets of PlGF and should play a decisive position in HREC organic features in DR.
Era of Uncooked RPPA Information and Their Conversion to Evaluation-Prepared Information.
Reverse section protein array (RPPA) supplies investigators with a strong high-throughput, quantitative, cost-effective know-how for purposeful proteomics research. It’s an antibody-based method with procedures much like that of Western blots. RPPA has all kinds of functions that vary from pharmacodynamics and drug sensitivity evaluation to biomarker discovery, subtype classification, and prediction of affected person prognosis and response to focused remedy. On this paper, we describe the know-how, its limitations, and a few options to beat them. We focus on the steps needed to acquire uncooked RPPA knowledge and convert them into strong, high-quality, analysis-ready knowledge.
We then illustrate the utility of the platform by highlighting some biomarkers and drug responses of most cancers cell strains that affirm earlier findings, as a method to validate the platform and the strategies offered right here. β-Amyloid protein (Aβ) putatively performs a seminal position in synaptic loss in Alzheimer’s illness (AD). Whereas there is no such thing as a consensus relating to the synaptic-relevant species of Aβ, it’s identified that Aβ oligomers (AβOs) are noticeably elevated within the early levels of AD, localizing at or throughout the synapse.
In cell and animal fashions, AβOs have been proven to connect to synapses and instigate synapse dysfunction and deterioration. To determine the pathological mechanism of synaptic loss in AD, will probably be necessary to determine the synaptic targets to which AβOs connect. Herein, we describe all of the steps required for the enrichment, isolation, and identification of SUMOylated substrates from human hepatoma cells and hepatic tissues from a liver most cancers mouse mannequin by utilizing SUBEs.
SUMO-Binding Entities (SUBEs) as Instruments for the Enrichment, Isolation, Identification, and Characterization of the SUMO Proteome in Liver Most cancers.
Submit-translational modification is a key mechanism regulating protein homeostasis and performance in eukaryotic cells. Amongst all ubiquitin-like proteins in liver most cancers, the modification by SUMO (Small Ubiquitin MOdifier) has been given probably the most consideration. Isolation of endogenous SUMOylated proteins in vivo is difficult as a result of presence of lively SUMO-specific proteases. Preliminary research of SUMOylation in vivo have been based mostly on the molecular detection of particular SUMOylated proteins (e.g., by western blot).
Nonetheless, in lots of instances, antibodies, usually made with non-modified recombinant protein, didn’t immunoprecipitate SUMOylated types of the protein of curiosity. Nickel chromatography has been the opposite strategy to check SUMOylation by capturing histidine-tagged variations of SUMO molecules. This strategy is especially utilized in cells stably expressing or transiently transfected with His-SUMO molecules. To beat these limitations, SUMO-binding entities (SUBEs) have been developed to isolate endogenous SUMOylated proteins.
 Aminoacetic acid, ?For electrophoresis |
|
GRM2233-100G |
EWC Diagnostics |
1 unit |
EUR 32.26 |
|
Description: Aminoacetic acid, ?For electrophoresis |
Aminoacetic acid, ?For electrophoresis |
|
GRM2233-500G |
EWC Diagnostics |
1 unit |
EUR 144.96 |
|
Description: Aminoacetic acid, ?For electrophoresis |
 Agarose, medium EEO ?For electrophoresis |
|
RM6249-100G |
EWC Diagnostics |
1 unit |
EUR 119.06 |
|
Description: Agarose, medium EEO ?For electrophoresis |
Agarose, medium EEO ?For electrophoresis |
|
RM6249-10G |
EWC Diagnostics |
1 unit |
EUR 19.63 |
|
Description: Agarose, medium EEO ?For electrophoresis |
Agarose, medium EEO ?For electrophoresis |
|
RM6249-1KG |
EWC Diagnostics |
1 unit |
EUR 1089.43 |
|
Description: Agarose, medium EEO ?For electrophoresis |
Agarose, medium EEO ?For electrophoresis |
|
RM6249-25G |
EWC Diagnostics |
1 unit |
EUR 36.87 |
|
Description: Agarose, medium EEO ?For electrophoresis |
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|
AJ846-017-175 |
Westburg |
each |
EUR 604.95 |
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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 70 |
 for electrophoresis, 99%) Ammonium Persulphate (APS) for electrophoresis, 99% |
|
28575 |
Sisco Laboratories |
25 Gms |
EUR 1.51 |
|
|
|
Description: Part A |
 ES 8/10 Double Gel Comb For ES Electrophoresis |
|
LE1011-A3 |
GenDepot |
Ea |
EUR 70 |
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|
20840057-1 |
Bio-WORLD |
500 mL |
EUR 91.56 |
Tris-HEPES-SDS Buffer 20X for Electrophoresis |
|
20840057-2 |
Bio-WORLD |
1 L |
EUR 173.27 |
 electrophoresis ps 1500v, 300ma, 150w |
|
EV2230 |
Consort |
ea |
EUR 898.8 |
electrophoresis ps 1500v, 300ma, 150w |
|
EV2230-UK |
Consort |
ea |
EUR 898.8 |
electrophoresis ps 1500v, 300ma, 150w |
|
EV2230-US |
Consort |
ea |
EUR 898.8 |
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|
EV2320 |
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EUR 898.8 |
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|
EV2320-UK |
Consort |
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EUR 898.8 |
electrophoresis ps 3000v, 150ma, 150w |
|
EV2320-US |
Consort |
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EUR 898.8 |
 ELECTROPHORESIS PS 1200V, 500MA, 300W |
|
EV3150 |
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ea |
EUR 1060 |
ELECTROPHORESIS PS 1200V, 500MA, 300W |
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EV3150-UK |
Consort |
ea |
EUR 1060 |
 ELECTROPHORESIS PS 3000V, 200MA, 300W |
|
EV3330 |
Consort |
ea |
EUR 1600 |
ELECTROPHORESIS PS 3000V, 200MA, 300W |
|
EV3330-UK |
Consort |
ea |
EUR 1600 |
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|
EV3620 |
Consort |
ea |
EUR 1600 |
ELECTROPHORESIS PS 6000V, 100MA, 300W |
|
EV3620-UK |
Consort |
ea |
EUR 1600 |
) Electrophoresis Buffer (10x) |
|
MBS2557002-100mL |
MyBiosource |
100mL |
EUR 85 |
Electrophoresis Buffer (10x) |
|
MBS2557002-200mL |
MyBiosource |
200mL |
EUR 90 |
Electrophoresis Buffer (10x) |
|
MBS2557002-500mL |
MyBiosource |
500mL |
EUR 95 |
Electrophoresis Buffer (10x) |
|
MBS2557002-5x500mL |
MyBiosource |
5x500mL |
EUR 380 |
 electrophoresis ps 400v, 500ma, 50w |
|
EV1450 |
Consort |
ea |
EUR 574.8 |
electrophoresis ps 400v, 500ma, 50w |
|
EV1450-UK |
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ea |
EUR 574.8 |
electrophoresis ps 400v, 500ma, 50w |
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EV1450-US |
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EUR 574.8 |
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EV2310 |
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EUR 750 |
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EUR 750 |
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|
EV2650 |
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ea |
EUR 750 |
ELECTROPHORESIS PS 600V, 500MA, 150W |
|
EV2650-UK |
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ea |
EUR 750 |
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|
EV3020 |
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ea |
EUR 1060 |
ELECTROPHORESIS PS 300V, 2000MA, 300W |
|
EV3020-UK |
Consort |
ea |
EUR 1060 |
 ELECTROPHORESIS PS 600V, 1000MA, 300W |
|
EV3610 |
Consort |
ea |
EUR 1060 |
ELECTROPHORESIS PS 600V, 1000MA, 300W |
|
EV3610-UK |
Consort |
ea |
EUR 1060 |
 TEMED, electrophoresis grade |
|
RM10358-100G |
EWC Diagnostics |
1 unit |
EUR 301.18 |
|
Description: TEMED, electrophoresis grade |
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|
EV245 |
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ea |
EUR 574.8 |
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|
LA690-1NO |
EWC Diagnostics |
1 unit |
EUR 432.84 |
|
Description: Electrophoresis Power Supply |
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|
21440026-1 |
Bio-WORLD |
15 mL |
EUR 21.21 |
Protein Antioxidant for SDS-PAGE Gel Electrophoresis |
|
21440026-2 |
Bio-WORLD |
25 mL |
EUR 28.49 |
Protein Antioxidant for SDS-PAGE Gel Electrophoresis |
|
21440026-3 |
Bio-WORLD |
50 mL |
EUR 43.44 |
Protein Antioxidant for SDS-PAGE Gel Electrophoresis |
|
21440026-4 |
Bio-WORLD |
100 mL |
EUR 77.25 |
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|
EV3150-US |
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EUR 1060 |
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|
EV3330-US |
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EUR 1600 |
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|
EV3620-US |
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ea |
EUR 1600 |
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|
MB570-10PR |
EWC Diagnostics |
1 unit |
EUR 38.47 |
|
Description: Agarose Gel Electrophoresis Kit |
 10X MOPS Electrophoresis Buffer |
|
ML050-500ML |
EWC Diagnostics |
1 unit |
EUR 32.7 |
|
Description: 10X MOPS Electrophoresis Buffer |
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|
EV2650-US |
Consort |
ea |
EUR 750 |
 CometAssay Electrophoresis System |
|
4250-050-ES |
Biotechne |
1 Kit |
EUR 3540.96 |
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|
EV2310-US |
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ea |
EUR 750 |
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|
EV3020-US |
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ea |
EUR 1060 |
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|
EV3610-US |
Consort |
ea |
EUR 1060 |
 ES Horizontal Electrophoresis Set |
|
LE1011 |
GenDepot |
Set |
EUR 410 |
 SDS-PAGE Electrophoresis Buffer |
|
MBS355536-1L |
MyBiosource |
1L |
EUR 165 |
SDS-PAGE Electrophoresis Buffer |
|
MBS355536-5x1L |
MyBiosource |
5x1L |
EUR 480 |
SDS-PAGE Electrophoresis Buffer |
|
MBS355542-10x1L |
MyBiosource |
10x1L |
EUR 250 |
 6X6 well Electrophoresis Apparatus |
|
EDK626 |
Bio Basic |
1 UNIT |
EUR 750.6 |
|
|
 electrophoresis power supply 120 vac |
|
EV245-US |
Consort |
ea |
EUR 574.8 |
 electrophoresis power supply uk plug |
|
EV245-UK |
Consort |
ea |
EUR 574.8 |
 myGel Mini Electrophoresis System, 115V |
|
BM0286 |
GenDepot |
System |
EUR 560 |
 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 |
|
Description: Alkaline Hemoglobin Electrophoresis Kit |
Alkaline Hemoglobin Electrophoresis Kit |
|
MBP008-224RUNS |
EWC Diagnostics |
1 unit |
EUR 208.42 |
|
Description: Alkaline Hemoglobin Electrophoresis Kit |
 Alkaline Paper Electrophoresis Kit (Cell |
|
MBP009-20RUNS |
EWC Diagnostics |
1 unit |
EUR 59.3 |
|
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 |
|
Description: Cellulose Acetate Electrophoresis System |
) Agarose Gel Electrophoresis Kit (Teaching) |
|
66602 |
Sisco Laboratories |
15 expt. Kit |
EUR 44.05 |
|
|
|
Description: Part E |
 ES Electrophoresis Casting Tray, 7cm X 7cm |
|
LE1011-A2 |
GenDepot |
Ea |
EUR 90 |
 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 |
|
|
|
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 100 |
 Hi-Eco Mini Horizontal Electrophoresis K |
|
LA851-1NO |
EWC Diagnostics |
1 unit |
EUR 205.56 |
|
Description: Hi-Eco Mini Horizontal Electrophoresis K |
) Electrophoresis Apparatus - 7x10cm (6-10-well) |
|
EDK611 |
Bio Basic |
1 UNIT |
EUR 383.11 |
|
|
) Electrophoresis Apparatus - 7x14cm (6-10-well) |
|
EDK613 |
Bio Basic |
1 UNIT |
EUR 487.51 |
|
|
 CometAssay Single Cell Gel Electrophoresis Assay |
|
4250-050-K |
Biotechne |
50 tests |
EUR 513.6 |
 ENDURO 7.7 Horizontal Electrophoresis 7 x 7cm Gels |
|
LE1001 |
GenDepot |
Set |
EUR 450 |
) Electrophoresis Apparatus - 7x7cm Dual (2x 6-10-well) |
|
EDK614 |
Bio Basic |
1 UNIT |
EUR 520.92 |
|
|
 ENDURO 7.10 Horizontal Electrophoresis 7 x 10cm Gels |
|
LE1002 |
GenDepot |
Set |
EUR 470 |
 ENDURO 10.10 Horizontal Electrophoresis 10 x 10cm Gels |
|
LE1003 |
GenDepot |
Set |
EUR 410 |
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|
LE1004 |
GenDepot |
Set |
EUR 610 |
 ENDURO 15.15 Horizontal Electrophoresis 15 x 15cm Gels |
|
LE1005 |
GenDepot |
Set |
EUR 710 |
 ENDURO 20.20 Horizontal Electrophoresis 20 x 20cm Gels |
|
LE1006 |
GenDepot |
Set |
EUR 750 |
 Gel Electrophoresis System with Time Control, 220-240 V |
|
Q-EP-2114 |
ABM |
1 |
EUR 1180 |
 10-well protein Vertical Electrophoresis Apparatus |
|
EDK692 |
Bio Basic |
1 UNIT |
EUR 487.51 |
|
|
 Gel Electrophoresis System with Muticaster, 100-120 V Input |
|
Q-EP-2034 |
ABM |
1 |
EUR 1715 |
 Gel Electrophoresis System with Muticaster, 220-240 V Input |
|
Q-EP-2056 |
ABM |
1 |
EUR 2125 |
 CometAssay Single Cell Gel Electrophoresis Assay-Silver |
|
4251-050-K |
Biotechne |
1 kit |
EUR 684.96 |
 Gel Electrophoresis System with Time Control, 100-120 V Input |
|
Q-EP-2100 |
ABM |
1 |
EUR 1385 |
) myGel Mini Electrophoresis System 230V (includes transformer) |
|
BM0291 |
GenDepot |
System |
EUR 580 |
myGel Mini Electrophoresis System 230V (includes transformer) |
|
E1101-E |
Benchmark Scientific |
1 PC |
EUR 665.55 |
 CometAssay Electrophoresis Starter Kit plus EU Power Supply |
|
4250-050-ESK-PS2 |
Biotechne |
1 Kit |
EUR 4597.68 |
 CometAssay Electrophoresis Starter Kit plus UK Power Supply |
|
4250-050-ESK-PS3 |
Biotechne |
1 Kit |
EUR 4597.68 |
 CometAssay Electrophoresis Starter Kit plus SZ Power Supply |
|
4250-050-ESK-PS4 |
Biotechne |
1 Kit |
EUR 4597.68 |
 NEW (specify 220V or 110V) $380) ComboMini? Electrophoresis System ? COMPLETE with ONE electrophoresis system + ONE gel maker set (4 combs, 1 large tray, 1 medium tray, 1 small tray, 1 gel casting stand) NEW (specify 220V or 110V) $380 |
|
ACT-CBMN-7 |
ACTGene |
each |
EUR 892.8 |
) SUPER Green II RNA gel stain *10,000× concentrate in DMSO* (Electrophoresis Grade) |
|
E37F007-100-uL |
EnoGene |
100 μL |
EUR 227 |
|
Description: Reagents for ssDNA or RNA electrophoresis |
SUPER Green II RNA gel stain *10,000× concentrate in DMSO* (Electrophoresis Grade) |
|
E37F007-50-uL |
EnoGene |
50 μL |
EUR 133 |
|
Description: Reagents for ssDNA or RNA electrophoresis |
) SUPER Green II RNA gel stain *10, 000× concentrate in DMSO* (Electrophoresis Grade) |
|
MBS8579833-005mL |
MyBiosource |
0.05mL |
EUR 195 |
SUPER Green II RNA gel stain *10, 000× concentrate in DMSO* (Electrophoresis Grade) |
|
MBS8579833-01mL |
MyBiosource |
0.1mL |
EUR 255 |
SUPER Green II RNA gel stain *10, 000× concentrate in DMSO* (Electrophoresis Grade) |
|
MBS8579833-5x005mL |
MyBiosource |
5x0.05mL |
EUR 820 |
SUPER Green II RNA gel stain *10, 000× concentrate in DMSO* (Electrophoresis Grade) |
|
MBS8579833-5x01mL |
MyBiosource |
5x0.1mL |
EUR 1100 |
 myGel InstaView™ Complete Electrophoresis System with Blue LED Illuminator |
|
E1201 |
Benchmark Scientific |
1 each |
EUR 1001.7 |
) SUPER Green I nucleic acid gel stain *10,000× concentrate in DMSO* (Electrophoresis Grade) |
|
E37F001-100-uL |
EnoGene |
100 μL |
EUR 227 |
|
Description: Reagents for dsDNA electrophoresis |
SUPER Green I nucleic acid gel stain *10,000× concentrate in DMSO* (Electrophoresis Grade) |
|
E37F001-500-uL |
EnoGene |
500 μL |
EUR 983 |
|
Description: Reagents for dsDNA electrophoresis |
SUPER Green I nucleic acid gel stain *10,000× concentrate in DMSO* (Electrophoresis Grade) |
|
E37F001-50uL |
EnoGene |
50 μL |
EUR 133 |
|
Description: Reagents for dsDNA electrophoresis |
) myGel Mini Electrophoresis System Starter Kit (Includes E1101, A1701 and W4000-100) |
|
E1101-SK |
Benchmark Scientific |
1 each |
EUR 727.06 |
) SUPER Green I nucleic acid gel stain *10, 000× concentrate in DMSO* (Electrophoresis Grade) |
|
MBS8579830-005mL |
MyBiosource |
0.05mL |
EUR 180 |
SUPER Green I nucleic acid gel stain *10, 000× concentrate in DMSO* (Electrophoresis Grade) |
|
MBS8579830-01mL |
MyBiosource |
0.1mL |
EUR 230 |
SUPER Green I nucleic acid gel stain *10, 000× concentrate in DMSO* (Electrophoresis Grade) |
|
MBS8579830-05mL |
MyBiosource |
0.5mL |
EUR 695 |
SUPER Green I nucleic acid gel stain *10, 000× concentrate in DMSO* (Electrophoresis Grade) |
|
MBS8579830-5x005mL |
MyBiosource |
5x0.05mL |
EUR 760 |
SUPER Green I nucleic acid gel stain *10, 000× concentrate in DMSO* (Electrophoresis Grade) |
|
MBS8579830-5x01mL |
MyBiosource |
5x0.1mL |
EUR 980 |
) High resolution protein SDS-PAGE electrophoresis pre mixed gel Kit (BIS Tris system) |
|
E1WP3231 |
EnoGene |
50T |
EUR 100 |
High resolution protein SDS-PAGE electrophoresis pre mixed gel Kit (BIS Tris system) |
|
E1WP3232 |
EnoGene |
100T |
EUR 150 |
) myGel Mini Electrophoresis System Starter Kit (Includes E1101-E, A1701 and W4000-100) |
|
E1101-SK-E |
Benchmark Scientific |
1 PC |
EUR 757.35 |
 myGel InstaView™ Complete Electrophoresis System with Blue LED Illuminator, 230V input |
|
E1201-E |
Benchmark Scientific |
1 each |
EUR 1058.9 |
Firstly, we describe strategies concerned within the preparation and lysis of the human hepatoma cells and liver tumor tissue samples. Then, an intensive rationalization of the preparation of SUBEs and controls is detailed together with the protocol for the protein pull-down assays. Lastly, some examples are supplied relating to the choices accessible for the identification and characterization of the SUMOylated proteome, particularly the usage of western–blot evaluation for the detection of a particular SUMOylated substrate from liver tumors or the usage of proteomics by mass spectrometry for high-throughput characterization of the SUMOylated proteome and interactome in hepatoma cells.
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