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Western QuickBlock Optimization Kit



Kit Description:

GenScript Western QuickBlock Kit and Western QuickBlock Optimization Kit are for quick blocking of any membranes used for Dot blot and Western blot. It can also be used for quick blocking of any solid surface such as microtiter plates used for ELISA. Instead of classical one-hour blocking of the solid surface with 5–10% dry milk, this kit enables the complete blocking of the solid surface in just 5 minutes. Furthermore, Western QuickBlock Kit significantly increases the Western detection sensitivity compared with dry milk blocking. And less amount of antibodies can be used to reach the same antigen detection level as using classical blocking method.

In some cases, due to the sequence similarity between an antigen and the blocking reagents, background from Western blot (also, Dot blot and ELISA) will be so high that the antigen cannot be detected clearly. To solve this problem, GenScript also introduced the Western QuickBlock Optimization Kits. With four different proprietary Western quick blot reagents provided in each kit, researchers can quickly find the best blocking reagents for any primary antibody in very short time.

The Western QuickBlock Kit and Western QuickBlock Optimization Kits are compatible with GenScript LumiSensorTM Chemiluminescent HRP Substrate Kit (L00221V60), LumiSensorTM Plus Chemiluminescent HRP Substrate Kit (L00225), and ChromoSensorTM One Solution TMB Substrate (L00222V60).

Key Features:

  • Quick blocking: The Western quickblock system takes only five minutes.
  • Quick optimization: finding the best blocking reagents in few hours instead of a few days.
  • Easy to perform: simple and quick procedure for blocking.
  • Sensitive: The Western quickblock makes your Western detection more sensitive.

Examples:

1. Shown below is a Western blot using the Western QuickBlock Kit (L00276)
Blocking using Western QuickBlock Kit was compared with the classical 10% dry milk blocking in Western blot detection of GAPDH protein from Hela cell lysate. Serially diluted Hela cell lysate samples were Western-blotted to WestClearTM nitrocellulose membrane after SDS-PAGE. The membrane was then cut into two halves and processed with the same procedures using Goat Anti-GAPDH Polyclonal Antibody (GenScript, A00191). The only difference is using different blocking reagents: classical 10% dry milk blocking (1.0 hour, left panel of Figure 1), and using Western QuickBlock Kit (5 min, right panel of Figure 1).


2. Shown below is a blot using the Western QuickBlock Optimization Kit (L00278)
Western QuickBlock Optimization Kit was used to optimize the Western blot detection of α-Tubulin in Hela cell lysate. 10 µg of Hela cell lysate was electrophoresised in 4 different lanes of SDS-PAGE and Western-blotted to WestClearTM nitrocellulose membrane. The membrane was then cut into four longitudinal strips and processed with the same procedures using unpurified monoclonal anti-α-Tubulin (Mouse Ascites Fluid, Sigma, T 5168). The only difference is using different blocking reagents as shown in Figure 2.

Technical Manual
  • TECHNICAL MANUAL: 3462_20071012094157.PDF (PDF)


  • Components
    Pretreat A-a solution 100 ml
    Pretreat A-b solution 100 ml
    Pretreat A-c solution 100 ml
    Pretreat A-d solution 100 ml
    Pretreat B 2 x 100 ml
    5-Slot Dot blot plate 1
    Protocol 1


    Document


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