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.

Agar, low gel strength

GK2227-250G 250 g
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Agar, low gel strength

GK2227-1KG 1 kg
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Agar, low gel strength

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Agar, high gel strength

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Agar, high gel strength

GK9085-1KG 1 kg
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Agar, high gel strength

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Agar, high gel strength

GK9085-500G 500 g
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Agar, low gel strength, suitable for microbiology

GC6234-100G 100 g
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Agar, low gel strength, suitable for microbiology

GC6234-1KG 1 kg
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Agar, low gel strength, suitable for microbiology

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Agar, low gel strength, suitable for microbiology

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Agarose, low EEO, GlenBiol, suitable for molecular biology

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Agarose, Low melting gel

AB0015 10g
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Agarose for DNA electrophoresis

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Agarose, medium EEO

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Agarose, medium EEO

GE4750-500G 500 g
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AGAROSE LE, LOW EEO, MOLECULAR BIOLOGY GRADE, 100G PER UNIT

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Description: Bioscience Mol Bio; Agarose

Amphotericin B, 5g

R1397-5g
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Apramycin Sulfate, 5g

R6571-5g
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5-FOA, 5g

R7039-5g
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ES 6 Tooth Gel Comb For ES Electrophoresis

LE1011-A4 Ea
EUR 124

ES 8/10 Double Gel Comb For ES Electrophoresis

LE1011-A3 Ea
EUR 124

CometAssay Single Cell Gel Electrophoresis Assay

4250-050-K 50 tests
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Silica Gel In Bag, 5G, 100/Pk

DSG111 1PK, 100UNIT
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AGAR, HIGH GEL

A01-102IHG-10kg 10 kg
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AGAR, HIGH GEL

A01-102IHG-2Kg 2 Kg
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AGAR, HIGH GEL

A01-102IHG-500g 500 g
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AGAROSE LE, LOW EEO, MOLECULAR BIOLOGY GRADE, 500G/UNIT, 10 UNITS PER CASE

AGR-LE-5001 1/pk
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Description: Bioscience Mol Bio; Agarose

Hi-Bind? T-Gel-Agarose

M1313-1
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Hi-Bind? T-Gel-Agarose

M1313-10
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Hi-Bind? T-Gel-Agarose

M1313-100
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CometAssay Single Cell Gel Electrophoresis Assay-Silver

4251-050-K 1 kit
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AGAROSE LE, LOW EEO, MOLECULAR BIOLOGY GRADE, 100G PER UNIT, 10 UNITS PER CASE

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Description: Bioscience Mol Bio; Agarose

HiQTM Low Melting Agarose

A0223-002 25g
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HiQTM Low Melting Agarose

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Agarose Gel DNA Extraction and Purification

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GelRed Agarose LE

41029-5G 5G
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GelGreen Agarose LE

41030-5G 5G
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Starch (Smithies), for electrophoresis

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Starch (Smithies), for electrophoresis

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Acrylamide, suitable for electrophoresis

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EUR 54

Agarose GPG/LMP low melt

A2501-100000 100 g
EUR 425
Description: Agarose has a wide range of physical, chemical and thermal stability, because of its greatest gelling ability that it acquires wide applications in biotechnology [2], such as agarose gel electrophoresis, protein purification, solid culture media, motility assays.

Agarose GPG/LMP low melt

A2501-25000 25 g
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Description: Agarose has a wide range of physical, chemical and thermal stability, because of its greatest gelling ability that it acquires wide applications in biotechnology [2], such as agarose gel electrophoresis, protein purification, solid culture media, motility assays.

Agarose II, Low Gelling Temperature

CH002 25 g
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Agarose II, Low Gelling Temperature

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Agarose gel DNA extraction and purification kit

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Preactivated Cys-Link-Affinity Gel Agarose (for Cys-containing peptides/proteins)

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E. Coli Proteins-Agarose affinity gel for removing E. coli antibodies

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Keyhole Limpet Hemocyanin (KLH)-Agarose affinity gel for removing KLH antibodies

KLH11-G 1 ml
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5G TPB

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HyAgarose? LM Agarose, Low Melting, 100g

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HyAgarose? LM Agarose, Low Melting, 10g

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HyAgarose? LM Agarose, Low Melting, 50g

R9012LM-50g
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5G Dithiothreitol (DTT)

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Keyhole Limpet Hemocyanin (KLH)-Agarose af is finity gel for removing KLH antibodies

KLH11-G-5 5 ml
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dam for 15cm gel

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G418 Disulfate (Low Endotoxin)

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Rabbit Anti-E. Coli Proteins IgG-Agarose affinity gel for removing E. coli proteins

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Cys-link agarose gel coupling buffer kit (5-10 peptides)

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Protein A-Agarose High-Binding 6% Highly Crosslinked

P9301-010 10ml
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Rabbit Gamma Globulin 5g

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gel casting base for evs3100

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gel casting base for evs3200

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Gel tray V (for Visirays)

2195934 2unit
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Description: Opt i on

Gel carrier for AE-6541

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Gellan Gum, Phyto Plant Culture Grade, ≥ 80% transmittance,Gel strength > 800 g/cm2

G2008-050 500g
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Gellan Gum, Phyto Plant Culture Grade, ≥ 80% transmittance,Gel strength > 800 g/cm2

G2008-100 1Kg
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electrophoresis power supply

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CometAssay Electrophoresis System

4250-050-ES 1 Kit
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Gellan Gum, Pure, Plant Culture Grade, ≥ 80% transmittance,Gel strength 400-700 g/cm2

G2007-050 500g
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Gellan Gum, Pure, Plant Culture Grade, ≥ 80% transmittance,Gel strength 400-700 g/cm2

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Gellan Gum, Pure, Plant Culture Grade, ≥ 80% transmittance,Gel strength 400-700 g/cm2

G2007-500 5Kg
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Agarose

20-abx186369
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  • 100 g
  • 25 g

Agarose

20-abx098147
  • EUR 217.00
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  • 100 g
  • 1 g

Agarose

abx082006-100g 100 g
EUR 244

Agarose

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Agarose

abx082008-100g 100 g
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G060-1 100 g
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Agarose

G060-2 500 g
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Agarose

Z5050007 500 g
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Agarose

Z5050013 100 g
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Maximo-Gel juice fluorescent Protein-Gel Gel stain

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Maximo-Gel juice fluorescent Protein-Gel Gel stain

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Rabbit THYMUS ACETONE POWDER 5g

41008-1 5 g
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supporting mask for midi gel dryer

EDRY1100-MASK ea
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ssilicone rubber for midi gel dryer

EDRY1100-SILRUB ea
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supporting mask for maxi gel dryer

EDRY1200-MASK ea
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silicone rubber for maxi gel dryer

EDRY1200-SILRUB ea
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Gel Casting S for WSE-1710

2322107 4unit
EUR 283
Description: Gel Tr ay (S) x4, Gel Cast i ng St and (S), S si zecomb (9wel l /5wel l ) x2

Gel Casting L for WSE-1710

2322108 4unit
EUR 283
Description: Gel Tr ay (L ) x2, Gel Cast i ng St and (L ), S si zecomb (22wel l /12wel l ) x2

Gel Casting set for 10.5x6cm gels

E1101-CS1 1 PC
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Gel Casting Stand for 5x6cm gels

E1101-CS2 1 PC
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Gel-Bright LED Gel Illuminator

E90003 1EA
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Description: Minimum order quantity: 1 unit of 1EA

6X6 well Electrophoresis Apparatus

EDK626 1 UNIT
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ES Horizontal Electrophoresis Set

LE1011 Set
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myGel Mini Electrophoresis System

BM0286 System
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CometAssay Electrophoresis Starter Kit

4250-050-ESK 1 Kit
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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

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Precast Agarose Gel TAE, 2%, 12 Well, EtBr Stained, 20 gels per Box

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Precast Agarose Gel TAE, 2%, 17 Well, EtBr Stained, 20 gels per Box

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Precast Agarose Gel TAE, 2%, 48 Well, EtBr Stained, 10 gels per Box

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Precast Agarose Gel TBE, 2%, 12 Well, EtBr Stained, 20 gels per Box

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Gel-FAST? Gel Staining/Destaining Kit

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TC Gel

CP048-005 500 g
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TC Gel

CP048-010 1 Kg
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Gel Tray

2394250 4unit
EUR 270
Description: I ncl udi ng pl at es

Gel Tray

2398194 2unit
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Gel Cutter

KS071012-11 12
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Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Gel Filter

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Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Casting Dams, For Enduro 7cm Gel System

LE1000CD-01 Ea
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Casting Dams, For Enduro 10cm Gel System

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Casting Dams, For Enduro 15cm Gel System

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Agarose StandardRoomTemperature

FYA002-100G 100 g Ask for price

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Agarose, Sepro

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Agarose, Sepro

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Agarose Tablets

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Agarose M

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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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