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Production of high-quality two-dimensional gel electrophoresis profile for marine medaka samples by using Trizol-based protein extraction approaches.

Production of high-quality two-dimensional gel electrophoresis profile for marine medaka samples by using Trizol-based protein extraction approaches.
Posted by Troy

Medaka Sea is one of the most popular models of fish species for Ecotoxicology and environmental research and study of proteomics is a useful tool for understanding the medaka molecular response after exposure to different environmental stresses. High-quality protein sample preparation is the key to producing high-quality two-dimensional gel electrophoresis (2-DE) results for proteomics analysis.

In recent years, the extraction of protein-based Trizol has gained popularity because of its performance promise in producing high-quality 2-DE and comfort method.Three Trizol-based approach (Trizol method, the method and the method of Trizol Trizol aliquots with commercial cleaning kit) is used to extract protein from marine samples medaka and 2-DE profiles produced. 2-DE profile quality and effectiveness of extraction methods were evaluated.

For comparison, two common protein extraction method (method of lysis buffer and trichloroacetic acid (TCA) / acetone precipitation extraction) are also applied in parallel to Trizol based approaches.Any three Trizol-based approach produces high-quality 2-DE profiles of marine medaka compared with both methods of lysis buffer and TCA / acetone precipitation extraction.

In addition, the Trizol method with clean-up kit produced the best commercial 2-DE profile in terms of the clarity of the background, the number of spots and resolution proteins.Trizol based approach offered a better choice than traditional protein extraction method for 2-DE analysis of medaka sea. A modified version of the method Trizol with commercial clean-up kit was shown to produce the best 2-DE profile.

Production of high-quality two-dimensional gel electrophoresis profile for marine medaka samples by using Trizol-based protein extraction approaches.
Production of high-quality two-dimensional gel electrophoresis profile for marine medaka samples by using Trizol-based protein extraction approaches.

A method of combining blue native polyacrylamide gel electrophoresis liquid chromatography-tandem mass spectrometry for the detection of circulating immune complexes between the monoclonal antibody therapy and anti-drug antibodies in animals.

Monoclonal antibody therapy can potentially induce unwanted immune response, so that the production of anti-drug antibodies (ADAS). Binding of fennel drug and subsequent formation of immune complexes (ICs) can trigger a variety of responses, depending on the size, concentration, and a subclass of fennel. To better understand the impact of fennel in pharmacokinetics, pharmacodynamics and toxicology profile, bioanalytical methods developed to detect IC between a human monoclonal immunoglobulin G (IgG) and fennel in a biological sample.

Regarding the experimental procedure, in brief, human-specific antibodies and human antibodies bound IC in biological samples separated by blue native polyacrylamide gel electrophoresis (BN-PAGE). The target fraction is then cut from the gel, followed by in-gel trypsin digestion and subsequent liquid chromatography-tandem mass spectrometry (LC-MS / MS) to monitor the human peptide-specific IgG. This method is capable of detecting various types of human antibodies with a lower detection limit of 10 ug / mL in monkey serum. Performance IC assay to detect the expressed using spiked samples, and pre-incubated in monkey serum IC clearly detectable.

ES Horizontal Electrophoresis Set

LE1011 Set
EUR 578.4

runVIEW horizontal electrophoresis system with 15 x 10cm gel tray 1 set of casting dams and 8 double sided combs

ELE2302 EACH
EUR 1681.2

runVIEW horizontal electrophoresis system 15 x 7cm gel tray 1 set of casting dams 8 double sided combs

ELE2300 EACH
EUR 1602

ES Electrophoresis Casting Tray, 7cm X 14cm

LE1011-A1 Ea
EUR 198

ES Electrophoresis Casting Tray, 7cm X 7cm

LE1011-A2 Ea
EUR 181.2

5 x SDS Electrophoresis Buffer

GR103039 1 L
EUR 107

4 x Phosphate/SDS Electrophoresis Buffer

GR103033 1 L
EUR 116

ES 8/10 Double Gel Comb For ES Electrophoresis

LE1011-A3 Ea
EUR 148.8

electrophoresis ps 1500v, 300ma, 150w

EV2230 ea
EUR 898.8

electrophoresis ps 1500v, 300ma, 150w

EV2230-UK ea
EUR 898.8

electrophoresis ps 1500v, 300ma, 150w

EV2230-US ea
EUR 898.8

electrophoresis ps 3000v, 150ma, 150w

EV2320 ea
EUR 898.8

electrophoresis ps 3000v, 150ma, 150w

EV2320-UK ea
EUR 898.8

electrophoresis ps 3000v, 150ma, 150w

EV2320-US ea
EUR 898.8

electrophoresis ps 1200v, 500ma, 300w

EV3150 ea
EUR 960

electrophoresis ps 1200v, 500ma, 300w

EV3150-UK ea
EUR 960

electrophoresis ps 1200v, 500ma, 300w

EV3150-US ea
EUR 960

electrophoresis ps 3000v, 300ma, 300w

EV3330 ea
EUR 1456.8

electrophoresis ps 3000v, 300ma, 300w

EV3330-UK ea
EUR 1456.8

electrophoresis ps 3000v, 300ma, 300w

EV3330-US ea
EUR 1456.8

electrophoresis ps 6000v, 150ma, 300w

EV3620 ea
EUR 1456.8

electrophoresis ps 6000v, 150ma, 300w

EV3620-UK ea
EUR 1456.8

electrophoresis ps 6000v, 150ma, 300w

EV3620-US ea
EUR 1456.8

electrophoresis ps 400v, 500ma, 50w

EV1450 ea
EUR 574.8

electrophoresis ps 400v, 500ma, 50w

EV1450-UK ea
EUR 574.8

electrophoresis ps 400v, 500ma, 50w

EV1450-US ea
EUR 574.8

electrophoresis ps 300v, 1000ma, 150w

EV2310 ea
EUR 686.4

electrophoresis ps 300v, 1000ma, 150w

EV2310-UK ea
EUR 686.4

electrophoresis ps 300v, 1000ma, 150w

EV2310-US ea
EUR 686.4

electrophoresis ps 600v, 500ma, 150w

EV2650 ea
EUR 686.4

electrophoresis ps 600v, 500ma, 150w

EV2650-UK ea
EUR 686.4

electrophoresis ps 600v, 500ma, 150w

EV2650-US ea
EUR 686.4

electrophoresis ps 300v, 2000ma, 300w

EV3020 ea
EUR 960

electrophoresis ps 300v, 2000ma, 300w

EV3020-UK ea
EUR 960

electrophoresis ps 300v, 2000ma, 300w

EV3020-US ea
EUR 960

electrophoresis ps 600v, 1000ma, 300w

EV3610 ea
EUR 960

electrophoresis ps 600v, 1000ma, 300w

EV3610-UK ea
EUR 960

electrophoresis ps 600v, 1000ma, 300w

EV3610-US ea
EUR 960

electrophoresis power supply

EV245 ea
EUR 574.8

ES 6 Tooth Gel Comb For ES Electrophoresis

LE1011-A4 Ea
EUR 148.8

miniONE Electrophoresis System

ELE3400 EACH
EUR 691.2

Agarose for DNA electrophoresis

16-0027-500 500 g
EUR 410.4

CometAssay Electrophoresis System

4250-050-ES 1 Kit
EUR 3540.96

Electrophoresis Apparatus - 7x10cm (6-10-well)

EDK611 1 UNIT
EUR 383.11

Electrophoresis Apparatus - 7x14cm (6-10-well)

EDK613 1 UNIT
EUR 487.51

myGel Mini Electrophoresis System

BM0286 System
EUR 741.6

myGel Mini Electrophoresis System

E1101 1 each
EUR 623.9

6X6 well Electrophoresis Apparatus

EDK626 1 UNIT
EUR 750.6

Chiller for Electrophoresis systems

CSLCHILLER EACH
EUR 6793.2

Starch (Smithies), for electrophoresis

GE6857-1KG 1 kg
EUR 370.8

Starch (Smithies), for electrophoresis

GE6857-500G 500 g
EUR 217.2

electrophoresis power supply 120 vac

EV245-US ea
EUR 574.8

CometAssay Single Cell Gel Electrophoresis Assay

4250-050-K 50 tests
EUR 513.6

electrophoresis power supply uk plug

EV245-UK ea
EUR 574.8

Gel Electrophoresis System with Time Control, 220-240 V

Q-EP-2114 1
EUR 1180

CometAssay Electrophoresis Starter Kit

4250-050-ESK 1 Kit
EUR 3749.28

Acrylamide, suitable for electrophoresis

GK5651-100G 100 g
EUR 64.8

Acrylamide, suitable for electrophoresis

GK5651-100 100
EUR 23.8

10-well protein Vertical Electrophoresis Apparatus

EDK692 1 UNIT
EUR 487.51

Electrophoresis Apparatus - 7x7cm Dual (2x 6-10-well)

EDK614 1 UNIT
EUR 520.92

Protein Antioxidant for SDS-PAGE Gel Electrophoresis

21440026-1 15 mL
EUR 17.3

Protein Antioxidant for SDS-PAGE Gel Electrophoresis

21440026-2 25 mL
EUR 23.23

Protein Antioxidant for SDS-PAGE Gel Electrophoresis

21440026-3 50 mL
EUR 35.42

Protein Antioxidant for SDS-PAGE Gel Electrophoresis

21440026-4 100 mL
EUR 62.99

Bovine Serum Albumin, 98% by electrophoresis

GE3699-10 10
EUR 28.4

Bovine Serum Albumin, 98% by electrophoresis

GE3699-100 100
EUR 142.4

Bovine Serum Albumin, 98% by electrophoresis

GE3699-25 25
EUR 50.6

Bovine Serum Albumin, 98% by electrophoresis

GE3699-250 250
EUR 284.8

Bovine Serum Albumin, 98% by electrophoresis

GE3699-500 500
EUR 506.2

Gel Electrophoresis System with Muticaster, 100-120 V Input

Q-EP-2034 1
EUR 1715

Gel Electrophoresis System with Muticaster, 220-240 V Input

Q-EP-2056 1
EUR 2125

Gel Electrophoresis System with Time Control, 100-120 V Input

Q-EP-2100 1
EUR 1385

CometAssay Single Cell Gel Electrophoresis Assay-Silver

4251-050-K 1 kit
EUR 684.96

Accuris myGel Mini Electrophoresis Syst w/ Transf

ELE1111 EACH
EUR 854.4

Tris-HEPES-SDS Buffer 20X for Electrophoresis

20840057-1 500 mL
EUR 74.66

Tris-HEPES-SDS Buffer 20X for Electrophoresis

20840057-2 1 L
EUR 139.51

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 each
EUR 892.8

myGel Mini Electrophoresis System 230V (includes transformer)

BM0291 System
EUR 789.6

myGel Mini Electrophoresis System 230V (includes transformer)

E1101-E 1 PC
EUR 652.5

CometAssay Electrophoresis Starter Kit plus EU Power Supply

4250-050-ESK-PS2 1 Kit
EUR 4597.68

CometAssay Electrophoresis Starter Kit plus UK Power Supply

4250-050-ESK-PS3 1 Kit
EUR 4597.68

CometAssay Electrophoresis Starter Kit plus SZ Power Supply

4250-050-ESK-PS4 1 Kit
EUR 4597.68

High resolution protein SDS-PAGE electrophoresis pre mixed gel Kit (BIS Tris system)

E1WP3231 50T
EUR 100

High resolution protein SDS-PAGE electrophoresis pre mixed gel Kit (BIS Tris system)

E1WP3232 100T
EUR 150

Low molecular weight protein SDS-PAGE electrophoresis pre mixed gel Kit (Tricine system)

E1WP326 50T
EUR 100

myGel InstaView™ Complete Electrophoresis System with Blue LED Illuminator

E1201 1 each
EUR 1001.7

High molecular weight protein SDS-PAGE electrophoresis pre mixed gel Kit (Tris acetate system)

E1WP324 50T
EUR 60

myGel Mini Electrophoresis System Starter Kit (Includes E1101, A1701 and W4000-100)

E1101-SK 1 each
EUR 712.8

OmniPage Mini Wide modular system inc tank capillary electrophoresis and electroblotting modules

VS10WCES EACH
EUR 1434.85

myGel Mini Electrophoresis System Starter Kit (Includes E1101-E, A1701 and W4000-100)

E1101-SK-E 1 PC
EUR 742.5

myGel InstaView™ Complete Electrophoresis System with Blue LED Illuminator, 230V input

E1201-E 1 each
EUR 1058.9

SUPER-Green-II-RNA-gel-stain-*10,000×-concentrate-in-DMSO*--(Electrophoresis-Grade)

E37F007 100-μL
EUR 226.67
Description: Reagents-for-ssDNA-or-RNA-electrophoresis

SUPER-Green-I-nucleic-acid-gel-stain-*10,000×-concentrate-in-DMSO*-(Electrophoresis-Grade)

E37F001 100-μL
EUR 226.67
Description: Reagents-for-dsDNA-electrophoresis

Axygen Horizontal Gel Box 20Cm

HGB-20 EACH
EUR 900

Horizontal head, for 12 x 1.5ml

BCM1604 1 pcs, 1 UNIT
EUR 200.68

Horizontal head, for 12 x 1.5ml

BV1000-H15 1 PC
EUR 108.5

Horizontal head for 12 x 1.5mL

MIX7018 EACH
EUR 146.4

Horizontal head, for 4 x 15ml

BCM1605 1 pcs, 1 UNIT
EUR 200.68

Horizontal head, for 2 x 50ml

BCM1606 1 pcs, 1 UNIT
EUR 200.68

Horizontal head, for 4 x 15ml

BV1000-H150 1 PC
EUR 108.5

Horizontal head, for 2 x 50ml

BV1000-H500 1 PC
EUR 108.5

Horizontal head for 4 x 15mL

MIX7020 EACH
EUR 146.4

Horizontal head for 2 x 50mL

MIX7022 EACH
EUR 146.4

AXYGEN® HORIZONTAL GEL BOX, 10 CM

HGB-10 1/pk
EUR 399.6
Description: Lab Equipment; Axygen Branded EQ

Midi Horizontal Gel Unit

ELE8120 EACH
EUR 630.42

6 x 15mL horizontal rotisserie 1 ea

MIX7080 EACH
EUR 92.4

6 x 50mL horizontal rotisserie 1 ea

MIX7082 EACH
EUR 92.4

6 x 15ml horizontal rotisserie, 1 ea

R4040-HZ15 1 PC
EUR 68

6 x 50ml horizontal rotisserie, 1 ea

R4040-HZ50 1 PC
EUR 68

HORIZONTAL TANK KIT

IB52200 -
EUR 55.88

AXYGEN® HORIZONTAL GEL BOX, 15 CM

HGB-15 1/pk
EUR 583.2
Description: Lab Equipment; Axygen Branded EQ

AXYGEN® HORIZONTAL GEL BOX, 7 CM

HGB-7 1/pk
EUR 343.2
Description: Lab Equipment; Axygen Branded EQ

Horizontal MicroLoops E

M8-L18SP-15H 20pcs.
EUR 196.6

Horizontal MicroLoops E

M8-L18SP-50H 20pcs.
EUR 196.6

TOOL REST, HORIZONTAL BAR

I4013 1EA
EUR 14.06

MAGic Clamp Tube Rack 4 x 15mL tubes horizontal

MIX7174 EACH
EUR 163.2

MAGic Clamp Tube Rack 2 x 50mL tubes horizontal

MIX7176 EACH
EUR 163.2

IBI QS710 HORIZONTAL UNIT

IB51000 7CM X 10CM FORMAT
EUR 509.62

IBI MP1015 HORIZONTAL UNIT

IB53000 10CM X 15CM FORMAT
EUR 675.93

IBI HR2025 HORIZONTAL UNIT

IB56000 20CM X 25CM FORMAT
EUR 877.32

IBI HR2525 HORIZONTAL UNIT

IB57000 25CM X 25CM FORMAT
EUR 965.82

Pipette Tray PVC Horizontal

PIP5500 EACH
EUR 44.4

Horizontal 50mL Tube Holder

MIX5088 EACH
EUR 176.4

MAGic Clamp Tube Rack 12 x 1.5/2.0mL tubes horizontal

MIX7172 EACH
EUR 163.2

MAGic Clamp™ Tube Rack, 4 x 15ml tubes, horizontal

H1000-MR-T150H 1 PC
EUR 122

MAGic Clamp™ Tube Rack, 2 x 50ml tubes, horizontal

H1000-MR-T500H 1 PC
EUR 122

mini horizontal unit 7x7 cm

EHS1101-SYS ea
EUR 381.6

midiplus horizontal unit 15x7-10-15 cm

EHS1300-SYS ea
EUR 496.8

mini horizontal unit 7x10 cm

EHS1102-SYS ea
EUR 381.6

midi horizontal unit 10x7 cm

EHS1201-SYS ea
EUR 422.4

mini horizontal unit 9x11 cm

EHS3200-SYS ea
EUR 595.2

wide horizontal unit 23.5x14 cm

EHS3600-SYS ea
EUR 930

wide horizontal unit 23.5x25 cm

EHS3610-SYS ea
EUR 1030.8

Horizontal orientator 384 wells

DD53423 PK2
EUR 78

midi horizontal unit 10x10 cm

EHS1202-SYS ea
EUR 422.4

maxi horizontal unit 20x10 cm

EHS1401-SYS ea
EUR 686.4

maxi horizontal unit 20x20 cm

EHS1402-SYS ea
EUR 727.2

maxi horizontal unit 20x25 cm

EHS1403-SYS ea
EUR 747.6

mini horizontal unit 12x14 cm

EHS3300-SYS ea
EUR 625.2

midi horizontal unit 15x15 cm

EHS3400-SYS ea
EUR 757.2

midi horizontal unit 15x25 cm

EHS3410-SYS ea
EUR 930

midi horizontal unit 20x25 cm

EHS3500-SYS ea
EUR 980.4

MAGic Clamp™ Tube Rack, 12 x 1.5/2.0ml tubes, horizontal

H1000-MR-T15H 1 PC
EUR 122

HS 260 Basic Horizontal Shaker

3066602 EACH
EUR 3482.4

midiplus horizontal unit 15x7 cm

EHS1301-SYS ea
EUR 433.2

INSTRUMENT REST, HORIZONTAL SLOTTED

I556 1EA
EUR 19.7

mini horizontal unit 7x7 & 7x10 cm

EHS1100-SYS ea
EUR 422.4

midiplus horizontal unit 15x10 cm

EHS1302-SYS ea
EUR 433.2

midiplus horizontal unit 15x15 cm

EHS1303-SYS ea
EUR 433.2

maxiplus horizontal unit 26x16 cm

EHS1501-SYS ea
EUR 939.6

maxiplus horizontal unit 26x24 cm

EHS1502-SYS ea
EUR 939.6

maxiplus horizontal unit 26x32 cm

EHS1503-SYS ea
EUR 939.6

Horizontal MicroLoops E - Assortment

M8-L18SP-HA1 20pcs.
EUR 196.6

midi horizontal unit 10x7 & 10x10 cm

EHS1200-SYS ea
EUR 453.6

Horizontal outer dims 415h x 140w x 560d with 11 apertures of approx 37mm

AH17911 EACH
EUR 184.8

Horizontal outer dims 415h x 140w x 560d with 6 apertures of approx 67mm

AH1796UT EACH
EUR 208.8

maxi horizontal unit 20x10 & 20x20 cm

EHS1400-SYS ea
EUR 727.2

wide horizontal unit 23.5x40 cm ex. port

EHS3620-SYS ea
EUR 1132.8

Horizontal Plastic Clip Microtube Holder

MIX5096 EACH
EUR 190.8

Tube Rack for 7x50ml QuEChERs, horizontal

BV1010-50HQ 1 PC
EUR 42.4

Tube Rack for 7x50mL QuEChERs horizontal

MIX7044 EACH
EUR 57.6

Genie Horizontal Orbital Shaker Attachments

MIX5321 EACH
EUR 603.6

maxiplus horizontal unit 26x16-24-32cm

EHS1500-SYS ea
EUR 1132.8

6 x 1.5/2.0ml tube holder, pack of 2 (Installed for vertical or horizontal mixing)

R4040-1520 1 PC
EUR 22.3

Casting Dams, For Enduro 10cm Gel System

LE1000CD-02 Ea
EUR 165.6

Horizontal shelf for freezer capacity for 16 boxes height 50 mm

DD37581 EACH
EUR 460.8

10 Well Mc Comb ,1mm Thick For Enduro 10cm Gel System

LE1000C10-01 Ea
EUR 153.6

10 Well Mc Comb ,1.5mm Thick For Enduro 10cm Gel System

LE1000C10-02 Ea
EUR 153.6

12 Well Comb ,1mm Thick For Enduro 10cm Gel System

LE1000C10-04 Ea
EUR 153.6

12 Well Comb ,1.5mm Thick For Enduro 10cm Gel System

LE1000C10-05 Ea
EUR 153.6

16 Well Comb ,1.5mm Thick For Enduro 10cm Gel System

LE1000C10-07 Ea
EUR 153.6

16 Well Comb ,2mm Thick For Enduro 10cm Gel System

LE1000C10-08 Ea
EUR 153.6

25 Well Comb ,1mm Thick For Enduro 10cm Gel System

LE1000C10-11 Ea
EUR 153.6

Taken together, these findings indicate that our method allows semi-quantitative analysis to estimate the ratio of human antibodies compared with the IC included in the total antibody. This method was successfully applied to the in vivo studies using rats, and the data helps explain the unexpectedly rapid clearance of human antibodies for the formation of large IC. The combination of IC separation by BN-PAGE and detection of peptide-specific human IgG with LC-MS / MS bioanalytical approach is common, useful for detecting IC in animals.

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