Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (4-12% gel, TGS buffer)
Sample
Mouse Tissue lysate - whole (Heart left ventricule)
Specification
Heart left ventricule
Treatment
Tunicamycin for 48h
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBST + 5% BSA
Dilution
1/1000
Secondary antibody
Name
Non-Abcam antibody was used: Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
ECL biorad Clarity
Bands
Specific: 140 kDa Non-specific: 200, 225 kDa
Exposure
30 second(s)
Positive control
Lane 3 and 4: Tunicamycin treatment for 48h showed an increase in P-IRE1 expression when compared to Sham (lane 1 and 2).
Additional data
Additional Notes
It might be useful to dilute the primary antibody at 1/1500 or 1/2000 to decrease the background noise (at 200-225 kDa).
Type of membrane: nitrocellulose (Biorad)
Lysis buffer: RIPA
Use of protease and phosphatase inhibitors (Roche)
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
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提交于 Dec 19 2014