Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Duodenum Biopsies)
Specification
Duodenum Biopsies
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
Other product details
Dilution
1/100
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 24°C · Diluent: TBS Tween20
Secondary antibody
Name
Non-Abcam antibody was used: chicken anti rabbit Alexa Fluor 488
Host species: Chicken
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Chicken
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Dilution
1/200
Additional data
Additional Notes
No CAMKII positive staining was observed in activated mucosal cells. We have previously use pERK to label activated mucosal cells in this tissue. Sections were negative, and looked the same as the negative control which had the primary antibody omitted. Different dilutions from 1/100 to 1/800 were trialled at varying temperatures overnight, as well as trialling PBSTx and TBSTween as wash buffers with no change in the result. TBST was also tested as a wash buffer with no change in staining. TBST resulted in much higher background fluoresence compared with PBST.
Abcam response
After we have investigated this case, Abcam decided the problem is due to a damaged vial. In this situation we offered a compensation because the product has been purchased in the last 120 days.
MRS. Nicole Isaacs
Verified customer
投稿 May 10 2010