Application
Western blot
Sample
Mouse Serum (serum (I used 10ul of 1:100 dilution of serum))
Gel Running Conditions
Reduced Denaturing (4-20% tris glycine)
Loading amount
1 µg
Specification
serum (I used 10ul of 1:100 dilution of serum)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C
Dilution
1/1000
Detection
Detection method
ECL
Exposure
10 second(s)
Bands
Specific: 30 kDa Non-specific: 52kda and 72kda kDa
Additional data
Additional Notes
I ran this in parallel and used antibody ab3455 and I got the correct bands from normal to experimental mice that matched 3 other ELISAs. The ab22554 gave me non-specific incorrect bands as well as bound adiponectin worse and did not show the difference between experimental and control mice.
Abcam response
Thank you for submitting this review. Adiponectin has a predicted size of 30 kD, but also several glycosylation sites which increase the MW of the protein. This may be why you see additional bands at ~52 and 72 kD. Our Abpromise guarantee does entitle you to a replacement antibody, credit or refund if the antibody is not working as it should and you have purchased within the past 12 months.
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
提交于 Oct 07 2015