GST-Tag Antikörper

Produktnummer ABIN5540371

Der Maus Monoklonal anti-GST-Tag Antikörper (Klon 3B2) (ABIN5540371) detektiert spezifisch GST-Tag in WB und IP. Dieser Antikörper reagiert spezifisch mit Proben aus Schistosoma japonicum.

Kurzübersicht für GST-Tag Antikörper (ABIN5540371)

Target: GST-Tag

Reaktivität: Schistosoma japonicum

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser GST-Tag Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunoprecipitation (IP)

Klon: 3B2

Produktdetails anti-GST-Tag Antikörper

Spezifität: This antibody recognizes recombinant GST-Tag specifically.

Aufreinigung: Protein A sepharose

Immunogen: Recombinant GST protein

Isotyp: IgG2b

Anwendungsinformationen

Applikationshinweise: Western blot: 1 μg/mL. Immunoprecipitation: 5 μg/100 μL of E. coli extracts containing GST - Tag fusion protein.

Protokoll: 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 suitable. 2) Resolve 10 μ l of sample by SDS - polyacrylamide gel electrophoresis (See the manufacture's manual for electrophoresis c onditions). 3) Transfer to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in a semi - dry transfer system. (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20 % MeOH). See the manufacture's manual for specific transfer procedure. 4) The transferred proteins can be visualized by staining the membrane for 1 minute with Ponceau. Rinse the membrane with PBS. 5) Non - specific binding sites are blocked by immersing the membrane in 5 % Skim Milk/PBS/0.05 % Tween 20 for 1 hr.at room temperat ure (20 - 25 o C) or for overnight at 4 o C. 6) Incubate in primary antibody diluted as suggested in the APPLICATIONS for 1 hour at room temperature(20 - 25 o C). (The optimal antibody concentration will depend on the experimental conditions.) 7) Wash the membrane 3 times with PBS containing 0.05 % Tween20 for 5~10 minutes each. 8) Incubate the membrane with secondary antibody (1:10,000 diluted horseradish peroxidase conjugated anti mouse IgG (H+L) antibody in PBS containing 0.05 % Tween 20 for 45 minutes at ro om temperature (20 - 25 o C). 9) Wash the membrane 3 times with PBS containing 0.05 % Tween20 for 5~10 minutes each. 10) Wipe excess buffer from the membrane, then incubate it with appropriate chemiluminescence reagents for 1 minute. Remove extra reagent from the membra ne by dabbing with a paper towel, and seal it in plastic wrap. 11) Expose to X - ray film in a dark room for 30 seconds. Develop the film as usual. The conditions for exposure and development may vary. Immunoprecipitation 1) Add 5 μ g of the antibody to 100 μ l of E. Coli extracts containing GST - Tag fusion protein. Mix well and incubate with gentle agitaiton for 30 - 120 minutes at 4 o C. 2) Add 20 μ l of 50 % Protein A - agarose beads. Mix well and incubate with gentle agitation for 1 hour at 4 o C. 3) Wash the beads 4 times with ice - cold Lysis buffer (50 mM HEPES, pH 7.3, 250 mM NaCl, 0.2 % N P - 40, 5 mM EDTA, 10 % glycerol), then centrifuge the tube at 2,500 x g for 10 seconds to remove the buffer. 4) Resuspend the beads in 40 μ l of Laemmli SDS PAGE sampl e buffer, boil for 3 - 5 minutes, and centrifuge for 5 minutes. Apply 20 μ l/lane for the SDS - PAGE analysis (See SDS - PAGE & Western blotting.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Format: Liquid

Buffer: PBS containing 50 % glycerol. No preservative is contained.

Konservierungsmittel: Azide free

Lagerung: -20 °C

Informationen zur Lagerung: Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: One year from despatch.

Details zu GST-Tag

Target: GST-Tag

Substanzklasse: Tag

Hintergrund: Expression vectors containing a protein and a tag protein are commonly used. GST - Tag fusion protein expression sys tem is preferably used in various laboratories, because its simple protein purification step by an affinity chromatograpy. This specific antibody for GST - Tag fusion protein is useful tools for monitoring of the fusion protein expression and affinity purifi cation.

UniProt: P08515