ONE-HOUR Western<sup>TM</sup> Advanced Kit (Mouse)

You are here: Products » Proteins, Expression, Isolation and Analysis » Protein Detection and Immunoassay » ONE-HOUR Western™ Detection Kits » ONE-HOUR Western^TM Advanced Kit » ONE-HOUR Western^TM Advanced Kit (Mouse)

ONE-HOUR Western^TM Advanced Kit (Mouse)

Kit Description:

GenScript's ONE-HOUR Western^TM Kit yields journal-quality Western or Dot blot results in about one hour. Using GenScript's breakthrough immunodetection technology (patent pending), the kit replaces the classical three-step western process, which can take nearly five hours. Transfer the proteins from gel to membrane and incubate it in the pretreat solution for five minutes. Then incubate in WB-2 solution with the mixture of primary antibody and WB-1 solution for 40 minutes. Then wash three times for five or ten minutes each. The membrane can then be developed with the HRP substrate. The ONE-HOUR Western^TM procedure is contrasted with a classical three-step Western at right.

The ONE-HOUR Western^TM Kit is designed to produce high signal with low background for quick and clear Western analysis of proteins.

Except for basic kits, all other kits contain all the necessary reagents, buffers and materials such as membrane for performing a Western blot, and no additional secondary antibody is needed.

The ONE-HOUR Western^TM Kit saves precious research time. For added cost-efficiency, we recommend our complete kits, which contain all the reagents, membrane, HRP substrate necessary for a perfect blot. However, the kit components are also sold separately.

Overview of ONE-HOUR Adcanced Western<sup>TM</sup> Procedure

Key Features:

Easy to perform: This kit has fewer and simpler steps than other Western kits.
Low background: The kit contains WestClear^TM Nitrocellulose Membrane and LumiSensor^TM series of Chemiluminescent HRP Substrate Kits optimized for low background.
Reproducible results: The kit produces highly reproducible results.
No additional secondary antibody is needed.

Storage:

Store WestClear^TM Nitrocellulose Membrane at room temperature. Store the rest of the kit at 4°C. It will remain stable for six months. Do not freeze the kit or any of its components.

Examples:

Comparison of the two ONE-HOUR Western^TM Blot Kits of different sensitivities using monoclonal antibodies:
Two similar blots were processed with the same procedures using different ONE-HOUR Western^TM Kits: Standard (L00205C) and Advanced (L00242). 10 µg and 2.5 µg of THE^TM Anti-GST Monoclonal Antibody (Mouse) (GenScript, A00865), respectively, were used with these two kits to detect GST protein.
The results are shown in Figure 1.

Figure 1. Western blots for the detection of GST protein by ONE-HOUR Western^TM using different ONE-HOUR Western^TM Kits: Standard (L00205C) and Advanced (L00242). 33.0, 11.0, 3.60, 1.20 0.41, 0.13, and 0.045 ng of GST protein were loaded into Lanes 1, 2, 3, 4, 5, 6, and 7 respectively.

Comparison of the two ONE-HOUR Western^TM Blot Kits of different sensitivities using polyclonal antibodies:
Two similar blots were processed with the same procedures using different ONE-HOUR Western^TM Kits: Standard (L00204C) and Advanced (L00241). 10 µg and 2.5 µg of Rabbit Anti-GST-tag Polyclonal Antibody (GenScript, A00097), respectively, were used with the two kits to detect GST protein.
The results are shown in Figure 2.

Figure 2. Western blots for the detection of GST protein by ONE-HOUR Western^TM using different ONE-HOUR Western^TM Kits: Standard (L00204C) and Advanced (L00241). 25.0, 12.5, 6.25, 3.10, 1.50, and 0.78 ng of GST protein were loaded into Lanes 1, 2, 3, 4, 5, and 6 respectively.

Technical Manual

Components

Application	Western detection (mouse)
Pretreat Solution A	50 ml
Pretreat Solution B	50 ml
WB-1 Solution	0.5 ml
WB-2 Solution	50 ml
5X Wash Solution	125 ml
WestClear^TM Nitrocellulose Membrane (0.2 μm, 7.5 X 8 cm)	5 sheets
LumiSensor^TM Chemiluminescent HRP Substrate^*	5 + 10 ml
User Manual	1

Document

Document-MSDS: 2333_20060818160706.PDF (PDF)

ONE-HOUR WesternTM Advanced Kit (Mouse)