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Plant Proteomics & Protein Expression

ProteoPrep

ProteoPrep Membrane Extraction Kit

ProteoPrep Total Extraction Sample Kit

ProteoPrep Universal Extraction Kit

 
ProteoPrep™ Membrane Extraction Kit
Description:

Sigma's ProteoPrep Membrane Extraction Kit is designed to prepare a highly enriched membrane protein solution from many types of cells. This kit also includes reagents for the reduction and alkylation of disulfide bonds. The cells are suspended in a low conductivity buffer and disrupted by ultrasonication or other appropriate method. The membranes and membrane proteins are centrifuged, washed with water and then resuspended in a chaotropic buffer solution. This protein solution is then reduced and alkylated. The end result is a soluble membrane protein sample that contains low salt and is ready for separation on isoelectric focusing (IEF), the first step in two dimensional gel electrophoresis. This kit contains enough reagents to generate at least six 2 ml samples.

 
Kit Components:
  • Soluble Cytoplasmic and Loosely-bound Membrane Protein Extraction Reagent, 3 bottles of powder, each reconstitutes to 125 ml
  • Cellular and Organelle Membrane Solubilizing Reagent, 1 bottle of powder that reconstitutes to a final volume of 23 ml
  • Tributylphosphine Stock Solution, 5 x 0.5 ml flame sealed ampules
  • Alkylating Reagent, Iodoacetamide, 5 x 56 mg in brown glass vials


Sample/Gel conditions
Source, E. coli,
1st dimension, IEF 4-7;
2nd dimension, 12% SDS-PAGE
Molloy, MP, et. al., Proteomic Analysis of the Escherichia Coli Outer Membrane, Eur. J. Biochem., 267, 2871-2881(2000).

Ordering Information

Product Product Name Package Size Technical Bulletin
PROT-MEM ProteoPrep Membrane Extraction Kit 1 Kit X

 
ProteoPrep Total Extraction Sample Kit
Description:

Sigma's ProteoPrep Sample Extraction Kit is designed to extract all of the proteins from any type of cell or tissue. It includes two reagents that have been used extensively for extraction and two new reagents with increased solubilizing power to extract more proteins. This kit also includes reagents for the reduction and alkylation of disulfide bonds. The cells are resuspended in any of the four chaotropic reagents provided and disrupted by ultrasonication or other appropriate method. The extract is clarified and the protein solution is then reduced and alkylated. The end result is a soluble total protein sample that contains low salt that is ready for separation on isoelectric focusing (IEF), the first step in two dimensional gel electrophoresis. This kit contains enough reagents to generate at least ten 2-ml samples using all four reagents.

 
Kit Components:
  • Chaotropic Membrane Extraction Reagent 1 (CHAPS/Urea), 1 bottle of powder that reconstitutes to a final volume of 25 ml
  • Chaotropic Membrane Extraction Reagent 2 (CHAPS/Urea/Thiourea/SB3-10), 1 bottle of powder that reconstitutes to a final volume of 25 ml
  • Chaotropic Membrane Extraction Reagent 3 (Proprietary formulation), 1 bottle of powder that reconstitutes to a final volume of 25 ml
  • Cellular and Organelle Membrane Solubilizing Reagent (Proprietary formulation), 1 bottle of powder that reconstitutes to a final volume of 23 ml
  • Tributylphosphine Stock Solution 5 x 0.5 ml flame sealed ampules
  • Alkylating Reagent, Iodoacetamide 5 x 56 mg in brown glass vials

Sample/Gel conditions
Source, E. coli, 1 mg protein load;
1st dimension, IEF 4-7;
2nd dimension, 12% SDS-PAGE
Herbert, B., Advances in protein solubilization for two-dimensional electrophoresis. Electrophoresis, 20, 660-663, (1999).

Ordering Information

Product Product Name Package Size Technical Bulletin
PROT-TOT ProteoPrep Total Extraction Sample Kit 1 Kit X

 
ProteoPrep Universal Extraction Kit
Description:

Sigma's ProteoPrep™ Universal Extraction Kit is designed to separate the soluble cytoplasmic and membrane proteins from many types of cells. This kit also includes reagents for the reduction and alkylation of disulfide bonds. The cells are resuspended in a low conductivity buffer and disrupted by ultrasonication or other appropriate method. The membranes and membrane proteins are centrifuged and then washed once with buffer. The soluble protein fraction is dried and then resuspended with a chaotropic buffer. The insoluble material is resuspended and solublized in another chaotropic buffer solution. Both protein solutions are then reduced and alkylated. The two resulting soluble protein samples contain low salts and are ready for separation by isoelectric focusing (IEF), the first step in two-dimensional gel electrophoresis. This kit contains reagents sufficient to generate ten or more 2-ml samples.

 
Kit Components:
  • Soluble Cytoplasmic Extraction Reagent, 2 bottles of powder, each reconstitutes to 125 ml of 5 mM Trizma®
  • Soluble Protein Resuspension Reagent, 1 bottle of powder that reconstitutes to a final volume of 23 ml of solution (pH 10.4) containing 7.0 M urea, 2.0 M thiourea, 1 % C7BzO detergent
  • Cellular and Organelle Membrane Solubilizing Reagent, 1 bottle of powder that reconstitutes to a final volume of 23 ml of solution (pH 10.4) containing 7.0 M urea, 2.0 M thiourea, 1 % C7BzO detergent, 40 mM Trizma
  • Tributylphosphine Stock Solution, 5 x 0.5 ml flame sealed ampules containing 200 mM tributylphosphine in N-methyl-2-pyrrolidone
  • Alkylating Reagent, Iodoacetamide, 5 x 56 mg in brown glass vials

Sample/Gel conditions
Source, E. coli
1st dimension, IEF 4-7;
2nd dimension, 12% SDS-PAGE

Ordering Information

Product Product Name Package Size Technical Bulletin
PROT-TWO ProteoPrep Universal Extraction Kit 1 Kit X