Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (human bladder transitional cell carcinoma)
Specification
human bladder transitional cell carcinoma
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5%
Other product details
Dilution
1/200
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 20°C
Secondary antibody
Name
Non-Abcam antibody was used: Dako HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Additional data
Additional Notes
1. How long was the sample fixation?
Overnight in Formaledehyde
2. Was a no primary antibody control at the same dilution carried out in parallel?
Control was pERK1/2 and pELK1
3. What buffer was used for antigen retrieval?
Citrate Buffer
4. What buffer was the antibodies diluted in?
5% BSA in PSA
5. Did you carry out permeabilization with Triton or Tween?
No
6. Did you try staining without antigen retrieval step?
No
7. Have you tried any optimization of the protocol?
Yes multiple concentrations with both TRIS/EDTA and Citrate buffer
Overnight in Formaledehyde
2. Was a no primary antibody control at the same dilution carried out in parallel?
Control was pERK1/2 and pELK1
3. What buffer was used for antigen retrieval?
Citrate Buffer
4. What buffer was the antibodies diluted in?
5% BSA in PSA
5. Did you carry out permeabilization with Triton or Tween?
No
6. Did you try staining without antigen retrieval step?
No
7. Have you tried any optimization of the protocol?
Yes multiple concentrations with both TRIS/EDTA and Citrate buffer
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. Imran Ahmad
Verified customer
提交于 Nov 29 2010