Application
Western blot
Sample
Rat Cell lysate - whole cell (cultured epithelial cells)
Loading amount
30 µg
Specification
cultured epithelial cells
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Other product details
Dilution
1/1000
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 22°C · Diluent: TTBS with 1% milk
Secondary antibody
Name
Non-Abcam antibody was used: Goat Anti mouse HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
Biorad WesternC, ImmunStar kit
Exposure
10 minute(s) and 0 second(s)
Positive control
We reblotted this membrane with a different anti-anexinV antibody (ab14196) and it did work. This antibody (ab54775) does not recognise annexinV in rat (but works great in human).
Additional data
Additional Notes
This antibody is very specific for human tissue - as stated in the specification sheet. In rat lysates we were unable to detect any AnnexinV bands with this antibody. For Rat, I would recommend the abcam Rabbit anti AnexinV antibody (ab14196).
Abcam response
Thank you for your review of ab54775 for staining western blots of rat cell lysates, and for your recommendation of ab14196 for this species. The immunogen for ab54775, recombinant human annexin V, is 92% identical to rat annexin V, but your data suggest that the antibody recognizes an epitope that is not conserved between the two species.
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.
Abcam user community
Verified customer
投稿 Oct 16 2012