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GST-Tag antibody (HRP)

Antigen

GST-Tag
Clonality Monoclonal (3B12)
Host
Alternatives

Mouse
Conjugate
Application
Alternatives Western Blotting (WB)
Catalog no. ABIN131911
Quantity 10 µl  (Variants)
Price Product not available in this region.
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Additional Information
Application Details Alternatives Variants back to top

Immunogen This antibody was purified from mouse ascites fluid using Protein-A Sepharose. This hybridoma was established by fusion of mouse myeloma cell SP2/0-Ag 14 with Balb/c mouse splenocyte immunized with recombinant GST protein.
Format Liquid
Isotype IgG2b
Clone 3B12
Description Expression vectors containing a protein and a tag protein are commonly used. The GST-Tag fusion protein expression system is preferably used in various laboratories, because of its simple protein purification step by affinity chromatography. This specific antibody for GST-Tag fusion protein is a useful tool for monitoring fusion protein expression and affinity purification.
Specificity This antibody recognizes recombinant GST-Tag specifically.

Application Details
Alternatives Variants back to top

Application Notes SDS PAGE & Western Blotting 1) Boil the sample for 3~5 minutes and centrifuge at 12,000 x g for a minute. Other methods may be employable. 2) Resolve 10 µl of sample by SDS- polyacrylamide gel electrophoresis. 3) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1mA/cm 2 for 1 hour in a semi-dry transfer system. (Transfer Buffer - 25mM Tris, 190mM glycine, 20% MeOH). See the manufacture's manual for the specific transfer procedure. 4) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH 7.4) for 1 hour at room temperature, or overnight at 4°C. 5) Incubate the membrane with the HRP conjugated anti-GST-Tag monoclonal antibody diluted with 1% skimmed milk (in PBS, pH 7.4) (1:1000) for 1 hour at room temperature. 6) Wash the membrane with PBS (5 minutes x 6 times). 7) Wipe excess buffer from the membrane, then incubate it with appropriate chemiluminescence reagents for 1 minute. Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap. 8) Expose to an X-ray film in a dark room for 5 minutes. Develop the film as usual. The conditions for exposure and development may vary. Western blotting: 1:1000 Detailed procedure is provided in the following Research Applications.
Buffer In PBS containing 1% BSA and 0.01% Benzalkonium Chloride.
Restrictions For Research Use only