Preliminary characterisation of two early meiotic wheat proteins after identification through 2D gel electrophoresis proteomics

Genetic-based approaches-including mutant screens population, microarray analysis, cloning and transgenesis have expanded our knowledge of gene function during meiosis in plants. Nonetheless, genetic tools are not without limitations. One alternative approach to studying meiosis plants, especially in polyploidy as Triticum aestivum L. (wheat bread), is proteomics. However, the proteins using proteomics-based approach is rarely depicted, with only two early learning efforts reported plant meiosis. Here, we report the initial investigation meiosis wheat bread using proteomics.

Five places differently expressed proteins were identified using 2D gel electrophoresis (2DGE) in the protein extracts of the four stages of meiosis were collected and three genotypes (wild-type Chinese Spring, ph1b and ph2a wheat mutant lines). mass spectrometry (MS / MS) identification tandem peptides from this protein led to the isolation and characterization of full-length clones of wheat Speckle-type POZ protein, a SF21-like protein and HSP70, and partial coding sequence of a hexose transporter.

Significantly, the function is suspected of Speckle-type POZ protein and HSP70 was confirmed using in vitro DNA binding test. Through the use of 2DGE proteomics approach, we show that proteomics is an alternative to the genetic-based approach when studying meiosis in wheat. More importantly, we reported a potential role for Speckle-type POZ protein and HSP70 in a chromosome pair during the early stages of meiosis in bread wheat.

Improved quantification of therapeutic proteins covalently aggregate by non-reducing capillary gel electrophoresis using sodium sulphate hexadecyl (CE-SHS)

Capillary gel electrophoresis (CGE) using sodium dodecyl sulphate (SDS-CGE or CE-SDS) is commonly used in the biotechnology industry to assess the purity of the complex therapy during the manufacturing process optimization and also for commercial release and stability testing. However, for therapeutic protein mAb-1 and mAb-2, non-reducing (NR) CE-SDS resulted in higher than expected% aggregates were much lower the purity obviously relative to the purity reported by complementary methods, such as Size Exclusion Chromatography (SEC ). In addition, a strong dependence of the protein load at the aggregate level was observed which prevents any reasonable assessment of the value of true purity.

The solution is to supplement the relatively hydrophobic SDS detergent hexadecyl sodium sulfate (SHS) in buffer gel sieving matrix protein that virtually eliminates the burden of dependency and reduce the value of the aggregate% to the level expected when compared to the SEC. Analytical ultracentrifugation (AUC) was used to help confirm the accuracy of the results of the SEC. This work underscores how to use detergents in addition to SDS in CGE applications can be valuable in commercial biological space and give examples of how the SEC can be used to confirm the accuracy of the data CGE.

 Preliminary characterisation of two early meiotic wheat proteins after identification through 2D gel electrophoresis proteomics
Preliminary characterisation of two early meiotic wheat proteins after identification through 2D gel electrophoresis proteomics

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

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.

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

CometAssay Single Cell Gel Electrophoresis Assay-Silver

4251-050-K 1 kit
EUR 684.96

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

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

E1WP324 50T
EUR 60

Acrylamide, suitable for electrophoresis

GK5651-100G 100 g
EUR 64.8

Acrylamide, suitable for electrophoresis

GK5651-100 100
EUR 23.8

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

E1WP326 50T
EUR 100

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

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

miniONE Electrophoresis System

ELE3400 EACH
EUR 691.2

5 x SDS Electrophoresis Buffer

GR103039 1 L
EUR 107

CometAssay Electrophoresis System

4250-050-ES 1 Kit
EUR 3540.96

myGel Mini Electrophoresis System

BM0286 System
EUR 741.6

ES Horizontal Electrophoresis Set

LE1011 Set
EUR 578.4

myGel Mini Electrophoresis System

E1101 1 each
EUR 623.9

6X6 well Electrophoresis Apparatus

EDK626 1 UNIT
EUR 750.6

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

electrophoresis power supply 120 vac

EV245-US ea
EUR 574.8

electrophoresis power supply uk plug

EV245-UK ea
EUR 574.8

ENDURO 7.7 Horizontal Electrophoresis 7 x 7cm Gels

LE1001 Set
EUR 651.6

CometAssay Electrophoresis Starter Kit

4250-050-ESK 1 Kit
EUR 3749.28

ENDURO 7.10 Horizontal Electrophoresis 7 x 10cm Gels

LE1002 Set
EUR 699.6

ENDURO 10.10 Horizontal Electrophoresis 10 x 10cm Gels

LE1003 Set
EUR 586.8

ENDURO 15.10 Horizontal Electrophoresis 15 x 10cm Gels

LE1004 Set
EUR 886.8

ENDURO 15.15 Horizontal Electrophoresis 15 x 15cm Gels

LE1005 Set
EUR 1081.2

ENDURO 20.20 Horizontal Electrophoresis 20 x 20cm Gels

LE1006 Set
EUR 1170

ES Electrophoresis Casting Tray, 7cm X 7cm

LE1011-A2 Ea
EUR 181.2

4 x Phosphate/SDS Electrophoresis Buffer

GR103033 1 L
EUR 116

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

ES Electrophoresis Casting Tray, 7cm X 14cm

LE1011-A1 Ea
EUR 198

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

EDK611 1 UNIT
EUR 383.11

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

EDK613 1 UNIT
EUR 487.51

Accuris myGel Mini Electrophoresis Syst w/ Transf

ELE1111 EACH
EUR 854.4

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

EDK614 1 UNIT
EUR 520.92

Agar, high gel strength

GK9085-100G 100 g
EUR 64.8

Agar, high gel strength

GK9085-1KG 1 kg
EUR 208.8

Agar, high gel strength

GK9085-250G 250 g
EUR 93.6

Agar, high gel strength

GK9085-500G 500 g
EUR 134.4

Agar, High Gel Strength

40100064-1 -
EUR 84.23

Agar, High Gel Strength

40100064-2 -
EUR 135.61

Agar, High Gel Strength

40100064-3 -
EUR 256.75

Agar, High Gel Strength

40100064-4 -
EUR 557.75

Agar, high gel strength

GK9085-1 1
EUR 142.4

Agar, high gel strength

GK9085-100 100
EUR 23.8

Agar, high gel strength

GK9085-250 250
EUR 47.4

Agar, high gel strength

GK9085-500 500
EUR 81.5

10-well protein Vertical Electrophoresis Apparatus

EDK692 1 UNIT
EUR 487.51

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

ELE2300 EACH
EUR 1602

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

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

Agar, low gel strength

GK2227-100G 100 g
EUR 60

Agar, low gel strength

GK2227-1KG 1 kg
EUR 170.4

Agar, low gel strength

GK2227-250G 250 g
EUR 79.2

Agar, low gel strength

GK2227-500G 500 g
EUR 112.8

Agar, low gel strength

GK2227-1 1
EUR 110.7

Agar, low gel strength

GK2227-100 100
EUR 19.8

Agar, low gel strength

GK2227-250 250
EUR 35.5

Agar, low gel strength

GK2227-500 500
EUR 63.3

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

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

myGel InstaView™ Complete Electrophoresis System with Blue LED Illuminator

E1201 1 each
EUR 1001.7

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

Agarose, High EEO

40100148-1 25 g
EUR 51.77

Agarose, High EEO

40100148-2 100 g
EUR 143.93

Agarose, high EEO

GE7747-1 1
EUR 660.6

Agarose, high EEO

GE7747-250 250
EUR 227.8

Agar, low gel strength, suitable for microbiology

GC6234-100G 100 g
EUR 64.8

Agar, low gel strength, suitable for microbiology

GC6234-1KG 1 kg
EUR 184.8

Agar, low gel strength, suitable for microbiology

GC6234-250G 250 g
EUR 84

Agar, low gel strength, suitable for microbiology

GC6234-500G 500 g
EUR 118.8

Agar, low gel strength, suitable for microbiology

GC6234-1 1
EUR 122.6

Agar, low gel strength, suitable for microbiology

GC6234-100 100
EUR 23.8

Agar, low gel strength, suitable for microbiology

GC6234-250 250
EUR 39.4

Agar, low gel strength, suitable for microbiology

GC6234-500 500
EUR 68.9

Agarose, low EEO

GE0001-100G 100 g
EUR 160.8

Agarose, low EEO

GE0001-1KG 1 kg
EUR 724.8

Agarose, low EEO

GE0001-250G 250 g
EUR 265.2

Agarose, low EEO

GE0001-25G 25 g
EUR 84

Agarose, low EEO

GE0001-500G 500 g
EUR 418.8

Agarose, low EEO

GE0001-5G 5 g
EUR 58.8

Agarose, Low EEO

40100152-1 50 g
EUR 75.92

Agarose, Low EEO

40100152-2 100 g
EUR 140.08

Agarose, Low EEO

40100152-3 500 g
EUR 446.01

Agarose, low EEO

GE0001-1 1
EUR 569.5

Agarose, low EEO

GE0001-100 100
EUR 102.7

Agarose, low EEO

GE0001-25 25
EUR 39.4

Agarose, low EEO

GE0001-250 250
EUR 189.8

Agarose, low EEO

GE0001-5 5
EUR 19

Agarose, low EEO

GE0001-500 500
EUR 316.4

Electrode Filling Sol Low Ionic Strength Applications

EFSLIAPP 100ML
EUR 101.84

Agarose LE Low EEO

AXY1000 PK10
EUR 2276.4

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.

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