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Protein Molecular Weight
Phosphorylase B 97,400
Bovine serum albumin 68,000 ...
OxyBlot™ Protein Standard
Description:
OxyBlot™ Protein Standard
Trade Name:
- OxyBlot
- Chemicon (Millipore)
Product Overview:
OxyBlot™ Protein Standard is composed of five proteins (phosphorylase B, BSA, ovalbumin, carbonic anhydrase, trypsin inhibitor) containing 1-3 dinitrophenylhydrazone (DNP) residues per protein. These attached DNP moieties permit the visualization of the proteins by the detection system of the OxyBlot™ Protein Oxidation Detection Kit.
Intended Use
OxyBlot™ Protein Standard serves as an internal control for the electrophoresis, western transfer and immunodetection steps of the OxyBlot™ procedure.
Intended Use
OxyBlot™ Protein Standard serves as an internal control for the electrophoresis, western transfer and immunodetection steps of the OxyBlot™ procedure.
Key Applications:
Western Blotting
Application Notes:
Material Provided:
Each vial contains 150 μl of OxyBlot™ Protein Standard, which is sufficient for 60 minigels.
Instructions for Use:
Warm the mixture to room temperature. (Since the mixture contains SDS that may precipitate during storage, it may be necessary to heat the stock solution to 50°C to redissolve the SDS). Combine 2.5 μl of the OxyBlot™ Protein Standard with 20 μl of 1X Gel Loading Buffer prior to loading onto the minigel. Do not heat prior to gel loading. Perform SDS-PAGE according to standard procedures.
NOTE: Since the proteins in the standard mixture already contain DNP residues, the protein standard should not be subjected to the derivatization reaction in the OxyBlot™ Protein Oxidation Detection Kit protocol.
In order to maintain consistency, the 2-Mercaptoethanol must be omitted from the 1X Gel Loading Buffer if a reducing agent was not used in the preparation of the experimental samples.
Reagent Preparation:
1X Gel Loading Buffer
To make 8 ml:
0.5 M Tris-HCl, pH 6.8 1.0 ml
Glycerol 0.8 ml
14.3 M 2-Mercaptoethanol (Reagent Grade) 0.1 ml
10% SDS 1.6 ml
0.05% Bromophenol Blue 0.32 ml
dH2O 4.18 ml
Divide into 1 ml aliquots and store at -15°C to -25°C.
Protein Molecular Weight
Phosphorylase B 97,400
Bovine serum albumin 68,000
Ovalbumin 43,000
Carbonic anhydrase 29,000
Trypsin inhibitor 21,000
OxyBlot™ is a trademark of Serologicals Company
Each vial contains 150 μl of OxyBlot™ Protein Standard, which is sufficient for 60 minigels.
Instructions for Use:
Warm the mixture to room temperature. (Since the mixture contains SDS that may precipitate during storage, it may be necessary to heat the stock solution to 50°C to redissolve the SDS). Combine 2.5 μl of the OxyBlot™ Protein Standard with 20 μl of 1X Gel Loading Buffer prior to loading onto the minigel. Do not heat prior to gel loading. Perform SDS-PAGE according to standard procedures.
NOTE: Since the proteins in the standard mixture already contain DNP residues, the protein standard should not be subjected to the derivatization reaction in the OxyBlot™ Protein Oxidation Detection Kit protocol.
In order to maintain consistency, the 2-Mercaptoethanol must be omitted from the 1X Gel Loading Buffer if a reducing agent was not used in the preparation of the experimental samples.
Reagent Preparation:
1X Gel Loading Buffer
To make 8 ml:
0.5 M Tris-HCl, pH 6.8 1.0 ml
Glycerol 0.8 ml
14.3 M 2-Mercaptoethanol (Reagent Grade) 0.1 ml
10% SDS 1.6 ml
0.05% Bromophenol Blue 0.32 ml
dH2O 4.18 ml
Divide into 1 ml aliquots and store at -15°C to -25°C.
Protein Molecular Weight
Phosphorylase B 97,400
Bovine serum albumin 68,000
Ovalbumin 43,000
Carbonic anhydrase 29,000
Trypsin inhibitor 21,000
OxyBlot™ is a trademark of Serologicals Company
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Kit or Assay Type:
Neurobiology Kits
Components:
Each vial contains 150 μl of OxyBlot Protein Standard, which is sufficient for 60 minigels.
Storage Conditions:
Prior to first use, store at 4°C. After using for the first time, divide the remaining mixture among several tubes. Store the aliquots at -15°C to -25°C. Dividing the mixture into aliquots serves to protect the mixture from excessive freeze-thaw cycles.