Application
Western blot
Sample
Mouse Tissue lysate - whole (Hippocampus)
Loading amount
20 µg
Specification
Hippocampus
Gel Running Conditions
Reduced Denaturing (4-20% polyacrylamide Bio-Rad ReadyGel, Tris buffer)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/3000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C
Secondary antibody
Name
Non-Abcam antibody was used: Cell Signaling #7074
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
Exposure
3 minute(s) and 0 second(s)
Bands
Specific: 43 kDa
Positive control
Mouse hippocampus lysate
Additional data
Additional Notes
We used a PVDF membrane, and we lysed the tissue in RIPA buffer.
The western blot was very clean, and we found that using lower concentrations of antibody yielded the positive band. We were able to drop out all non-specific bands by tweaking the exposure conditions, indicating that the positive band is the strongest. To us, this is the best indicator of whether an antibody can be reliably used for immunohistochemistry.
The western blot was very clean, and we found that using lower concentrations of antibody yielded the positive band. We were able to drop out all non-specific bands by tweaking the exposure conditions, indicating that the positive band is the strongest. To us, this is the best indicator of whether an antibody can be reliably used for immunohistochemistry.
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.
Mr. Michael Saul
Verified customer
提交于 Aug 11 2011