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°C.
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：10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information：0.45 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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