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Tau are stabilising proteins for axonal microtubules within the Central Nervous System, and as such are also recognised as Mictrotubule-associated Protein (MAP). Tau is predominantly active in the distal portions of axons to carry out its function.
1. HOW DID OUR ANTIBODY PERFORM? CUSTOMER REVIEW: Tau Polyclonal Antibody (STJ95911)
2. WHAT WAS USED? Primary Antibody: Provider: Dilution ratio: Secondary Antibody: Application: Materials for validation: STJ95911 Tau Antibody St John’s Laboratory 1:500 Goat Anti-Rabbit IgG Antibody, (H+L) HRP conjugate (Sigma) Western Blot. Murine intestinal longitudinal muscle myenteric plexus (LMMP)
3. WHAT WAS THE PROTOCOL? Treatment of materials: LMMP were homogenized using a polytron tissue grinder in RIPA buffer (150 mM NaCl, 50 mM Tris-HCl, 0.25% wt/vol sodium deoxycholate, 0.1% Nonidet P- 40, 100 M NaVO4) with protease inhibitors (Sigma). Samples were treated with loading buffer for 5 min in water bath at 100¡ãC. Gel electrophoresis: 10% polyacrylamide gel, constant voltage 160 V, 60 min. Transfer: 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V, 90 min. Blocking: 1X TBST with 5% bovine serum albumin for 30 min. Primary antibody probing: primary antibody was added to the solution of 1X TBST with 5% bovine serum albumin, 4?C for overnight. Membrane wash: 1X TBST washes for 3 times. Secondary antibody probing: secondary antibody was added to the solution of 1X TBST with 5% bovine serum albumin with 1:2000 dilution ratio, for 45 min. Membrane wash: 1X TBST washes for 3 times. Visualization: ECL visualization for 5 min. Images were captured using Hyper Film MP (GE Healthcare).
4. WHAT WAS THE PROTOCOL? Gel electrophoresis information£º10% polyacrylamide gel, constant voltage 160 V, 60 min. Transfer information£º0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 90 min. No. Antigen Loading amount Primary antibody Primary antibody dilution ratio Secondary antibody dilution ratio Target band KD Visualization time 1 LMMP from C57Bl6J control mouse 20 ?g STJ95911 Tau Antibody 1:500 1:2000 ~70KD 60S 2 LMMP from C57Bl6J mouse infected with neurotropic virus
5. WHAT DID THE CUSTOMER THINK? Testing results were provided by Ignazio Castagliuolo from University of Padova, Italy. He stated that ¡°In LMMP isolated from both control and infected C57Bl6J mice, there were target bands detected by STJ95911 Tau Antibody.¡±
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