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One-Step Western^TM Advanced Kit (Rabbit)

Kit Description:

GenScript's One-Step WesternTM 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-Step WesternTM procedure is contrasted with a classical three-step Western at right. The One-Step WesternTM 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 and X-ray films for performing a Western blot, and no additional secondary antibody is needed. The One-Step WesternTM Kit saves precious research time. For added cost-efficiency, we recommend our complete kits, which contain all the reagents, membrane, HRP substrate, and X-ray film necessary for a perfect blot. However, the kit components are also sold separately.

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, and X-ray film, 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.

Order:

Cat.No. Name Size Price Quantity   L00241 One-Step WesternTM Advanced Kit (Rabbit)  1 Kit (10 Assays) $ 345.00

Technical Manual: Download PDF file TM0352 Components: Components L00241 L00242 L00243 Application  Western detection (rabbit)  Western detection (mouse)  Western detection (goat)  Pretreat Solution A  100 ml  100 ml  100 ml  Pretreat Solution B  100 ml  100 ml  100 ml  WB-1 Solution  1 ml  1 ml  1 ml  WB-2 Solution  100 ml  100 ml  100 ml  10X Wash Solution  125 ml  125 ml  125 ml  WestClearTM Nitrocellulose Membrane (0.2 µm, 7.5 x 8 cm)  10 sheets  10 sheets  10 sheets  LumiSensorTM Chemiluminescent HRP Substrate*  10 + 20 ml  10 + 20 ml  10 + 20 ml  High-Sensitivity X-Ray Film (5 x 7 inches)  20 sheets  20 sheets  20 sheets  Protocol  1  1  1  PROTOCOL:  20080716033645  (PDF) Examples: Comparison of the three One-Step WesternTM Blot Kits of different sensitivities using monoclonal antibodies: Two similar blots were processed with the same procedures using different One-Step WesternTM Kits: Standard (L00205C) and Advanced (L00242). 10 µg and 2.5 µg of THETM 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-Step WesternTM using different One-Step WesternTM 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 three One-Step WesternTM Blot Kits of different sensitivities using polyclonal antibodies: Two similar blots were processed with the same procedures using different One-Step WesternTM 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-Step WesternTM using different One-Step WesternTM 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.

Related Products:   Code Name Size Price Quantity L00242  One-Step WesternTM Advanced Kit (Mouse)  1 Kit (10 Assays)  $ 345.00 L00243  One-Step WesternTM Advanced Kit (Goat)  1 Kit (10 Assays)  $ 345.00 L00354  LumiSensorTM Super HRP Substrate  1 kit  $ 135.00 L00358  High-Sensitivity X-Ray Film (5 X 7 inches)  100 sheets  $ 65.00 L00358  High-Sensitivity X-Ray Film (5 X 7 inches)  20 sheets   $ 14.90