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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 482

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

LE1000C10-10 Ea
EUR 128

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

LE1000C10-01 Ea
EUR 128

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

LE1000C10-02 Ea
EUR 128

Casting Dams, For Enduro 10cm Gel System

LE1000CD-02 Ea
EUR 138

AXYGEN® HORIZONTAL GEL BOX, 10 CM

HGB-10 1/pk
EUR 333
Description: Lab Equipment; Axygen Branded EQ

99445-10 DCT 10 X 75MM

99445-10 250/pk
EUR 63
Description: Disposable Culture Tubes; DCT's, CGW

6 x 15ml horizontal rotisserie

R4040-HZ15 1 PC
EUR 106.47

6 x 50ml horizontal rotisserie

R4040-HZ50 1 PC
EUR 164.69

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

LE1000C10-03 Ea
EUR 128

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

LE1000C10-04 Ea
EUR 128

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

LE1000C10-05 Ea
EUR 128

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

LE1000C10-06 Ea
EUR 128

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

LE1000C10-07 Ea
EUR 128

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

LE1000C10-08 Ea
EUR 128

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

LE1000C10-11 Ea
EUR 128

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

LE1000C10-12 Ea
EUR 128

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

LE1000C10-09 Ea
EUR 128

Horizontal head, for 12 x 1.5ml

BCM1604 1 pcs, 1 UNIT
EUR 167.23

Horizontal head, for 4 x 15ml

BCM1605 1 pcs, 1 UNIT
EUR 167.23

Horizontal head, for 2 x 50ml

BCM1606 1 pcs, 1 UNIT
EUR 167.23

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

EDK611 1 UNIT
EUR 319.26

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

EDK613 1 UNIT
EUR 406.26

10-well protein Vertical Electrophoresis Apparatus

EDK692 1 UNIT
EUR 406.26

Nitrocellulose Membrane, 0.25 µm, 9 cm x 10cm

AR0135-02 20 sheets/Pack
EUR 96

Nitrocellulose Membrane, 0.45 µm, 9 cm x 10cm

AR0135-04 20 sheets/Pack
EUR 96

ES Electrophoresis Casting Tray, 7cm X 14cm

LE1011-A1 Ea
EUR 165

ES Electrophoresis Casting Tray, 7cm X 7cm

LE1011-A2 Ea
EUR 151

midiplus horizontal unit 15x7-10-15 cm

EHS1300-SYS ea
EUR 414

Horizontal head

BV1000-H15 1 PC
EUR 134.38

Horizontal head

BV1000-H150 1 PC
EUR 134.38

Horizontal head

BV1000-H500 1 PC
EUR 2344.4

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

EDK614 1 UNIT
EUR 434.1

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

GELS Polyclonal Antibody

ES11007-100ul 100ul
EUR 279
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 207
Description: A Rabbit Polyclonal antibody against GELS from Human/Mouse/Rat. This antibody is tested and validated for WB, ELISA, WB, ELISA

Anti-GELS antibody

STJ192165 200 µl
EUR 197
Description: Unconjugated Rabbit polyclonal to GELS

GELS Polyclonal Antibody

ABP58629-003ml 0.03ml
EUR 158
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 289
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 414
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

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

HGB10-10-GT 1/pk
EUR 74
Description: Lab Equipment; Axygen Branded EQ

Gel Tray 10 X 10 Cm

LE1000GT-03 Ea
EUR 142

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 99.28
Description: Western Blot Boxes

ES 8/10 Double Gel Comb For ES Electrophoresis

LE1011-A3 Ea
EUR 124

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

LE1000C15-10 Ea
EUR 131

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

LE1000C20-10 Ea
EUR 138

Daunomycin (10.10), DNA Aptamer, FITC labeled

AD-113-F Custom Ask for price

ENDURO™ PowerStack

LE0304S Ea
EUR 604

electrophoresis power supply

EV245 ea
EUR 479

CometAssay Electrophoresis System

4250-050-ES 1 Kit
EUR 2950.8

Tenacissoside X

N2110-10 10 mg
EUR 572
Description: Extracted from Marsdenia tenacissima (Roxb.) Wight et Arn.;Store the product in sealed, cool and dry condition

X-NeuNAc

A3944-10 10 mg
EUR 131
Description: Km: 0.89 mM for neuraminidase X-Neu5Ac is a new substrate for chromogenic assay of neuraminidase activity in bacterial expression systems.

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

LE1000C15-01 Ea
EUR 131

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

LE1000C15-02 Ea
EUR 131

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

LE1000C7-01 Ea
EUR 124

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

LE1000C7-02 Ea
EUR 124

Kodak X-Ray Film 8 x 10 inch

B503 100 Sheets
EUR 202

1730 10 SNAP-SEAL 10 OZ

1730-10 100/pk
EUR 80
Description: Disposable Plastic; Plastic Containers

SC-10

B6299-10 10 mg
EUR 213

GSA 10

B5767-10 10 mg
EUR 229

MRT 10

B7685-10 10 mg
EUR 238

6X6 well Electrophoresis Apparatus

EDK626 1 UNIT
EUR 625.5

Starch (Smithies), for electrophoresis

GE6857-1KG 1 kg
EUR 309

Starch (Smithies), for electrophoresis

GE6857-500G 500 g
EUR 181

Acrylamide, suitable for electrophoresis

GK5651-100G 100 g
EUR 54

myGel Mini Electrophoresis System

BM0286 System
EUR 618

Agarose for DNA electrophoresis

16-0027-500 500 g
EUR 342

CometAssay Electrophoresis Starter Kit

4250-050-ESK 1 Kit
EUR 3124.4

MultiBox for Western blotting, 4 compartments, 32 x 52 x 28mm each (11/4 x 2 5/16 x 11/8 in.), for various gels

B1203 6/pack
EUR 95.9
Description: Western Blot Boxes

PSB 10 hydrochloride

B6923-10 10 mg
EUR 366

10-DEBC hydrochloride

B7105-10 10 mg
EUR 221
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.

MFZ 10-7

B5595-10 10 mg
EUR 350

TC ASK 10

B5728-10 10 mg
EUR 438

KinaSorb™ 10

KE785-10 10 preps
EUR 461

AI-10-49

A8694-10 10 mg
EUR 212
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 492
Description: Mouse monoclonal Keratin 10 antibody

Human Factor X (Factor 10) Antibody

12212-05011 150 ug
EUR 217

Gel Tray 7 X 10 Cm

LE1000GT-02 Ea
EUR 134

Gel Tray 15 X 10 Cm

LE1000GT-04 Ea
EUR 151

10 x TBE (Tris/Borate/EDTA)

A00269 500ml
EUR 56.36

Triton ® X-114 (10%) Solution

B1015-100
EUR 126

Triton ® X-114 (10%) Solution

B1015-500
EUR 185

APF LB Miller, 10 x 1L

0133-S 1L each
EUR 105

mini horizontal unit 7x7 cm

EHS1101-SYS ea
EUR 318

mini horizontal unit 7x10 cm

EHS1102-SYS ea
EUR 318

midi horizontal unit 10x7 cm

EHS1201-SYS ea
EUR 352

midi horizontal unit 10x10 cm

EHS1202-SYS ea
EUR 352

midiplus horizontal unit 15x7 cm

EHS1301-SYS ea
EUR 361

midiplus horizontal unit 15x10 cm

EHS1302-SYS ea
EUR 361

midiplus horizontal unit 15x15 cm

EHS1303-SYS ea
EUR 361

maxi horizontal unit 20x10 cm

EHS1401-SYS ea
EUR 572

maxi horizontal unit 20x20 cm

EHS1402-SYS ea
EUR 606

maxi horizontal unit 20x25 cm

EHS1403-SYS ea
EUR 623

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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