Application
Western blot
Sample
Rat Tissue lysate - other (hippocampus)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10%)
Loading amount
30 µg
Specification
hippocampus
Blocking step
Milk as blocking agent for 5 hour(s) and 0 minute(s) · Concentration: 7% · Temperature: 25°C
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C
Dilution
1/5000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: anti rabbit HRP-linked IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ecl
Exposure
5 minute(s) and 0 second(s)
Additional data
Additional Notes
We followed your protocol for western blot research on normal rat hippocampus. The protocol said the diluted concentration could be 1:5000-1:20000. However,we diluted this antibody in 1:4000 getting no bond of any protein. Our lysis buffer contained pmsf. Our membrane was pvdf . Our control such as beta-actin was normal indicating that our manual was okay. We stored your product at -20.
Abcam response
Abcam is currently re-testing this antibody for reactivity with rat samples by western blot. The reviewer's sample and gel-runing conditions were non-reduced and non-denatured. The antibody has stained rat samples well by ICC but there is a possiblity that the epitope the antibody recognizes is more available in reduced/denatured samples than in native.
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. House Liu
Verified customer
投稿 Nov 07 2014