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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.

5 gel caster for 8 x 10cm or 10 x 10cm gels.

CSL6CAST EACH
EUR 508.44

ES Horizontal Electrophoresis Set

LE1011 Set
EUR 578.4

Dummy Plate 10 x 10cm

VS10DP EACH
EUR 23.94

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

LE1000C10-10 Ea
EUR 153.6

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

AXYGEN® HORIZONTAL GEL BOX, 10 CM

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

Casting Dams, For Enduro 10cm Gel System

LE1000CD-02 Ea
EUR 165.6

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

ELE2302 EACH
EUR 1520.76

Multi Sub Midi 10 x 10cm UV Tray

MS10UV10 EACH
EUR 79.8

Vert Sub Mini 10 x 10cm Dual Caster

VS10DCAST EACH
EUR 196.08

Replacement Rubber mats for 10 x 10cm caster

VS10DCASTM EACH
EUR 50.16

Burnsoothe Burn Relief Dressing 10cm x 10cm

SAF1294 PK10
EUR 47.88

CoolTherm burn relief dressing 10cm x 10cm

SAF1330 PK10
EUR 164.16

99445-10 DCT 10 X 75MM

99445-10 250/pk
EUR 75.6
Description: Disposable Culture Tubes; DCT's, CGW

6 x 15ml horizontal rotisserie

R4040-HZ15 1 PC
EUR 127.76

6 x 50ml horizontal rotisserie

R4040-HZ50 1 PC
EUR 197.63

5 x SDS Electrophoresis Buffer

GR103039 1 L
EUR 128.4

C18 3um 10cm x 3mm

CHR4804 EACH
EUR 644.1

AQ 3um 10cm x 2.1mm

CHR4990 EACH
EUR 664.62

AQ 5um 10cm x 4.6mm

CHR4992 EACH
EUR 607.62

GC18 4um 10cm x 4.6mm

CHR5004 EACH
EUR 579.12

Dummy Plate 20 x 10cm

VS10WDP EACH
EUR 26.22

12 Well Comb ,0.75mm Thick For Enduro 10cm Gel System

LE1000C10-03 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 ,0.75mm Thick For Enduro 10cm Gel System

LE1000C10-06 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

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

LE1000C10-12 Ea
EUR 153.6

10 x 10cm Notched Glass Plates 2mm thick Pack of 2

VS10NG PK2
EUR 38.76

Horizontal head, for 12 x 1.5ml

BCM1604 1 pcs, 1 UNIT
EUR 200.68

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 12 x 1.5mL

MIX7018 EACH
EUR 121.98

Horizontal head for 4 x 15mL

MIX7020 EACH
EUR 121.98

Horizontal head for 2 x 50mL

MIX7022 EACH
EUR 121.98

4 x Phosphate/SDS Electrophoresis Buffer

GR103033 1 L
EUR 139.2

SU C18 3.5u 10cm x 3.0mm

CHR4376 EACH
EUR 687.35

C18 5um 10cm x 30mm id

CHR4826 EACH
EUR 4822.2

C18 Polar 3u 10cm x 2.1mm

CHR4864 EACH
EUR 643.61

MSP-C4 5um 10cm x 2.1mm

CHR5014 EACH
EUR 593.62

MSP-C4 5um 10cm x 3.0mm

CHR5016 EACH
EUR 593.62

MSP-C4 5um 10cm x 4.0mm

CHR5018 EACH
EUR 593.62

MDC-C18 5um 10cm x 3.0mm

CHR5048 EACH
EUR 612.36

MDC-C18 5um 10cm x 4.0mm

CHR5050 EACH
EUR 612.36

MDC-C18 5um 10cm x 4.6mm

CHR5052 EACH
EUR 612.36

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

EDK611 1 UNIT
EUR 383.11

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

EDK613 1 UNIT
EUR 487.51

10-well protein Vertical Electrophoresis Apparatus

EDK692 1 UNIT
EUR 487.51

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

LE1000C10-09 Ea
EUR 153.6

Kinesis Tubing PEEKsil 1/32 x 25um x 10cm

CHR2453 PK2
EUR 43.32

Kinesis TuBING PEEKsil 1/32 x 50um x 10cm

CHR2463 PK2
EUR 43.32

Kinesis Tubing PEEKsil 1/16 x 25um x 10cm

CHR2593 PK5
EUR 110.58

Kinesis Tubing PEEKsil 1/16 x 50um x 10cm

CHR2611 PK5
EUR 102.6

Kinesis Tub SS 1/32 x 205um x 10cm

CHR4823 EACH
EUR 13.68

Kinesis Tub SS 1/32 x .015 x 10cm

CHR4841 EACH
EUR 20.52

6 x 15mL horizontal rotisserie 1 ea

MIX7080 EACH
EUR 76.38

6 x 50mL horizontal rotisserie 1 ea

MIX7082 EACH
EUR 76.38

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

CoolTherm burn relief dressing 10cm x 40cm

SAF1332 PK7
EUR 152.76

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

ELE2300 EACH
EUR 1448.94

midiplus horizontal unit 15x7-10-15 cm

EHS1300-SYS ea
EUR 496.8

Genorise® 10 x RNA Sample Loading Buffer

GR102010-10 10 mL
EUR 177.6

Nitrocellulose Membrane, 0.25 µm, 9 cm x 10cm

AR0135-02 20 sheets/Pack
EUR 115.2

Nitrocellulose Membrane, 0.45 µm, 9 cm x 10cm

AR0135-04 20 sheets/Pack
EUR 115.2

Multi Sub Choice 15 x 10cm UV Tray

MS15UV10 EACH
EUR 93.48

Multi Sub Maxi 20 x 10cm UV Tray

MS20UV10 EACH
EUR 124.26

Multi Sub Mini 7 x 10cm UV Tray

MS7UV10 EACH
EUR 72.96

LU C18 (2) 3um 10cm x 3mm id

CHR3964 EACH
EUR 643.61

Horizontal head

BV1000-H15 1 PC
EUR 161.26

Horizontal head

BV1000-H150 1 PC
EUR 161.26

Horizontal head

BV1000-H500 1 PC
EUR 2813.28

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

EDK614 1 UNIT
EUR 520.92

Daunomycin (10.10), DNA Aptamer, Biotinylated

AD-113-B Custom Ask for price

Daunomycin (10.10), DNA Aptamer, unlabeled

AD-113-U Custom Ask for price

Technical Buffer pH 10.10 250ml

51350010 EACH
EUR 25.99

MAGic Clamp Tube Rack 4 x 15mL tubes horizontal

MIX7174 EACH
EUR 133.38

MAGic Clamp Tube Rack 2 x 50mL tubes horizontal

MIX7176 EACH
EUR 133.38

Kinesis Tubing SS 1/16od x 0.02id - 10cm Length

CHR4789 EACH
EUR 5.7

Kinesis Tubing SS 1/32 x 0.018 - 10cm Length

CHR4849 EACH
EUR 20.52

Kinesis Tubing SS 1/16od x 0.03id - 10cm Length

CHR4855 EACH
EUR 7.98

Kinesis Tubing SS 1/16 x .040 - 10cm Length

CHR4887 EACH
EUR 10.26

Kinesis Tubing SS 1/16 x .046 - 10cm Length

CHR4901 EACH
EUR 11.4

AXYGEN® GEL TRAY 10 X 10 CM FOR USE WITH 10 CM GEL BOX, UV TRANSPARENT

HGB10-10-GT 1/pk
EUR 88.8
Description: Lab Equipment; Axygen Branded EQ

ES 8/10 Double Gel Comb For ES Electrophoresis

LE1011-A3 Ea
EUR 148.8

GELS Polyclonal Antibody

ES11007-100ul 100ul
EUR 334.8
Description: A Rabbit Polyclonal antibody against GELS from Human/Mouse/Rat. This antibody is tested and validated for WB, ELISA, WB, ELISA

GELS Polyclonal Antibody

ES11007-50ul 50ul
EUR 248.4
Description: A Rabbit Polyclonal antibody against GELS from Human/Mouse/Rat. This antibody is tested and validated for WB, ELISA, WB, ELISA

GELS Polyclonal Antibody

ABP58629-003ml 0.03ml
EUR 189.6
Description: A polyclonal antibody for detection of GELS from Human, Mouse, Rat. This GELS antibody is for WB, ELISA. It is affinity-purified from rabbit serum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from part region of human GELS protein

GELS Polyclonal Antibody

ABP58629-01ml 0.1ml
EUR 346.8
Description: A polyclonal antibody for detection of GELS from Human, Mouse, Rat. This GELS antibody is for WB, ELISA. It is affinity-purified from rabbit serum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from part region of human GELS protein

GELS Polyclonal Antibody

ABP58629-02ml 0.2ml
EUR 496.8
Description: A polyclonal antibody for detection of GELS from Human, Mouse, Rat. This GELS antibody is for WB, ELISA. It is affinity-purified from rabbit serum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from part region of human GELS protein

Anti-GELS antibody

STJ192165 200 µl
EUR 236.4
Description: Unconjugated Rabbit polyclonal to GELS

20 Well Comb ,0.75mm Thick For Enduro 15cm Gel System

LE1000C15-10 Ea
EUR 157.2

50 Well Comb ,2mm Thick For Enduro 20cm Gel System

LE1000C20-10 Ea
EUR 165.6

Gel Tray 10 X 10 Cm

LE1000GT-03 Ea
EUR 170.4

ENDURO™ PowerStack

LE0304S Ea
EUR 724.8

Horizontal MicroLoops E

M8-L18SP-15H 20 MOUNTS
EUR 112
Description: Horizontal MicroLoops E; box of 20 each 15x150 um aperture mounted on 18 mm rods

Horizontal MicroLoops E

M8-L18SP-50H 20 MOUNTS
EUR 112
Description: Horizontal MicroLoops E; box of 20 each 50x500 um aperture mounted on 18 mm rods

Daunomycin (10.10), DNA Aptamer, FITC labeled

AD-113-F Custom Ask for price

electrophoresis power supply

EV245 ea
EUR 574.8

CometAssay Electrophoresis System

4250-050-ES 1 Kit
EUR 3540.96

miniONE Electrophoresis System

ELE3400 EACH
EUR 624.72

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

MIX7172 EACH
EUR 133.38

TBE Bufer, 10 x

GR103009 1 L
EUR 99.6

TBST Bufer, 10 x

GR103010 1 L
EUR 128.4

TE Bufer, 10 x

GR103012 500 mL
EUR 190.8

Silicon Septa x 10

WAT4826 PK10
EUR 144.78

MultiBox for Western blotting, 4 compartments, 38 x 71 x 28mm each (11/2 x 2 13/16 x 11/8 in.), for various gels including half-mini protein gels

B1204 6/pack
EUR 119.14
Description: Western Blot Boxes

Kinesis Tubing SS 1/32 x 250um (0.010) - 10cm Length

CHR4831 EACH
EUR 21.66

Kinesis Tubing SS 1/16 x 180um(.007in) - 10cm Length

CHR4863 EACH
EUR 6.84

Kinesis Tubing SS 1/16 x 125um(.005in) - 10cm Length

CHR4915 EACH
EUR 6.84

9 x 10cm Notched Glass Plates with 0.75mm Bonded Spacers

VS10NGS075 EACH
EUR 53.58

9 x 10cm Plain Glass Plates with 0.75mm Bonded Spacers

VS10PGS075 EACH
EUR 35.34

19 x 10cm Notched Glass Plates with 0.75mm Bonded Spacers

VS10WNGS075 EACH
EUR 125.4

19 x 10cm Notched Glass Plates with 1mm Bonded Spacers

VS10WNGS1 EACH
EUR 132.24

Accuris Gel Casting Set for 5x6cm Gels (18/10 Teeth)

ELE1114 EACH
EUR 86.64

10 Well Comb ,1mm Thick For Enduro 15cm Gel System

LE1000C15-01 Ea
EUR 157.2

10 Well Comb ,1.5mm Thick For Enduro 15cm Gel System

LE1000C15-02 Ea
EUR 157.2

10 Well Comb , 1mm Thick For Enduro 7cm Gel System

LE1000C7-01 Ea
EUR 148.8

10 Well Comb , 1.5mm Thick For Enduro 7cm Gel System

LE1000C7-02 Ea
EUR 148.8

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

AH17911 EACH
EUR 149.34

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

AH1796UT EACH
EUR 168.72

X-NeuNAc

A3944-10 10 mg
EUR 157.2
Description: Km: 0.89 mM for neuraminidase X-Neu5Ac is a new substrate for chromogenic assay of neuraminidase activity in bacterial expression systems.

Tenacissoside X

N2110-10 10 mg
EUR 686.4
Description: Extracted from Marsdenia tenacissima (Roxb.) Wight et Arn.;Store the product in sealed, cool and dry condition

Platform for 10 x 10 mL tubes

MIX1256 EACH
EUR 47.88

Digital Hotplate, 10 x 10 inch, 115V

H3710-H 1 each
EUR 545.3

Digital Hotplate, 10 x 10 inch, 230V

H3710-H-E 1 each
EUR 545.3

Magnetic Stirrer, 10 x 10 inch, 115V

H3710-S 1 each
EUR 545.3

Magnetic Stirrer, 10 x 10 inch, 230V

H3710-SE 1 each
EUR 545.3

DIGE gel 3 gels

28937451 EACH
EUR 486.78

1730 10 SNAP-SEAL 10 OZ

1730-10 100/pk
EUR 96
Description: Disposable Plastic; Plastic Containers

9 x 10cm Plain Glass Plates 2mm thick Pack of 2

VS10PG EACH
EUR 28.5

GSA 10

B5767-10 10 mg
EUR 274.8

SC-10

B6299-10 10 mg
EUR 255.6

MRT 10

B7685-10 10 mg
EUR 285.6

Kodak X-Ray Film 8 x 10 inch

B503 100 Sheets
EUR 242.4

Horizontal 50mL Tube Holder

MIX5088 EACH
EUR 144.78

Pipette Tray PVC Horizontal

PIP5500 EACH
EUR 38.76

Midi Horizontal Gel Unit

ELE8120 EACH
EUR 630.42

Horizontal orientator 384 wells

DD53423 PK2
EUR 63.84

10 x Cell Lysis Buffer

GR103003x10 200 mL
EUR 141.6

10 x PBS, pH 7.30

GR103004x10 1 L
EUR 100.8

10 x Universal Blocking Buffer

GR103005 200 mL
EUR 154.8

10 x ELISA Wash Buffer

GR103014 1 L
EUR 117.6

10 x ELISA Reagent Diluent

GR103028 100 mL
EUR 92.4

Hypercassette Neutral 10 X 12In

RPN11650 EACH
EUR 367.08

4 X 10 CARBON CARTRIDGE

CDFC410F2 EACH
EUR 295.26

Bellows pasteur pipette x 10

WAT4772 PK10
EUR 144.78

CometAssay Electrophoresis Starter Kit

4250-050-ESK 1 Kit
EUR 3749.28

Agarose for DNA electrophoresis

16-0027-500 500 g
EUR 410.4

myGel Mini Electrophoresis System

BM0286 System
EUR 741.6

Acrylamide, suitable for electrophoresis

GK5651-100G 100 g
EUR 64.8

Starch (Smithies), for electrophoresis

GE6857-1KG 1 kg
EUR 370.8

Starch (Smithies), for electrophoresis

GE6857-500G 500 g
EUR 217.2

6X6 well Electrophoresis Apparatus

EDK626 1 UNIT
EUR 750.6

10cm UV Gel Scoop

MS10UVS EACH
EUR 51.3

Chiller for Electrophoresis systems

CSLCHILLER EACH
EUR 6453.54

myGel Mini Electrophoresis System

E1101 1 each
EUR 623.9

Kinesis Adapt - 10-32M x 10-32F - PEEK

CHR4283 EACH
EUR 27.36

AI-10-49

A8694-10 10 mg
EUR 254.4
Description: AI-10-49 is a selective inhibitor of CBF? -SMMHC and RUNX1 interaction with a FRET IC50 value of 260nM. AI-10-49 restores RUNX1 transcriptional activity, displays favorable pharmacokinetics, and delays leukemia progression in mice.

Keratin 10 antibody

10-2541 250 ug
EUR 590.4
Description: Mouse monoclonal Keratin 10 antibody

MFZ 10-7

B5595-10 10 mg
EUR 420

TC ASK 10

B5728-10 10 mg
EUR 525.6

PSB 10 hydrochloride

B6923-10 10 mg
EUR 439.2

10-DEBC hydrochloride

B7105-10 10 mg
EUR 265.2
Description: 10-DEBC hydrochloride is a selective inhibitor of Akt (or termed PKB) [1], with an IC50 value of approximate 48 ?M [2].Akt is a type of serine/threonine kinase. It phosphorylates and inactivates components in the apoptotic machinery, including Caspase 9 and BAD.

KinaSorb™ 10

KE785-10 10 preps
EUR 553.2

Empty Tubes (10)

2MLET-10 10
EUR 17
Description: Empty tubes used for preparation of urine samples for use with the Oxalate Quick Test Strips. Quantity: 10 empty 2mL tubes.

Aluminum Test Tube Basket 10 x 6 x 6

HS20341B EACH
EUR 40.47

Finntip 10 0.2-10ul Pink 10 x 96 rack

PIP3644 PK960
EUR 111.72

Cytiva ALUMINA PLATES 10 X 8 CM 10 EA

GESE202N10 PK10
EUR 327.18

Aluminium Tray For 10 X 10 Wide Vials 28.5mm

FLY1320 EACH
EUR 93.48

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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