Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rhesus monkey Tissue sections (ovary)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Reveal Decloacker (Biocare Medical)
Permeabilization
No
Specification
ovary
Blocking step
Avidin/Biotin blocking kit (Vector) then TBS + 10% goat serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Modified Davidson's Fixative (EMS)
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBS-0.3% Triton X-100
Dilution
1/100
Secondary antibody
Dilution
1/200
Name
Non-Abcam antibody was used: Vectastain Elite ABC Kit (Rabbit IgG) #PK-6101
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Additional data
Additional Notes
The ovary of a rhesus monkey was fixed in Modified Davidson’s Fixative (EMS) and paraffin-embedded. 5µm sections were deparaffinized in Citrisolv (Fisher) and antigen retrieval achieved using Reveal Decloaker (Biocare Medical). Sections were prepared for immunohistochemistry using Avidin/Biotin blocking kit (Vector Labs) in 10% normal goat serum in TBS. Sections were then incubated with diluted 1:100 HMGB1 primary antibody (ab18256, Abcam) overnight at 4°C. Sections were washed and then incubated with diluted 1:200 biotinylated goat anti-rabbit secondary antibody from the Vectastain Elite ABC kit (Vector Labs) for 2 hours at room temperature, washed again, and followed by a 30 minute incubation in the ABC reagent (Vectastain Elite ABC kit Vector Labs) and a final wash. Binding was detected with diaminobenzidine (DAB Vector Labs) for 6 minutes. Image of sections incubated with HMGB1 antibody + HMGB1 blocking peptide (Abcam #ab18650) can also be provided.
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.
Mrs. Lindsay Reustle
Verified customer
提交于 May 27 2015