Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (RPE-1)
Specification
RPE-1
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton X100 in PBS
Other product details
Dilution
1/200
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 23°C · Diluent: PBS
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-Rabbit Cy3
Host species: Goat
Clonality: Polyclonal
Conjugation: Cy3 ®
Host species: Goat
Clonality: Polyclonal
Conjugation: Cy3 ®
Dilution
1/200
Additional data
Additional Notes
Asynchronous RPE-1 cells were paraformaldehyde fixed (4%, 10min), permeablized and immunofluorescently labeled. Cells were incubated with primary antibody for 1hr or overnight at both 1:100 and 1:200 dilutions. Subsequent testing was also performed on methanol fixed cells with similar conditions. None of these fixation/labeling protocols appear to be compatible as only background staining was evident. In fact, the accompanying image has an exposure time that is so long for the SMG5 channel, that the background has become evident/grey.
Abcam response
Thank you for your valuable feedback. ICC/IF was not a tested application for ab33033 and it seems that this antibody does not recognize SMG5 under these experimental conditions. Thank you for your time - we hope that this information will be helpful for future customers.
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.
DR. Kirk Mcmanus
Verified customer
提交于 Sep 04 2007