Sample
Human Tissue sections (Normal Skin and Basal Cell Carcinoma/Solar Elastos)
Application
Immunohistochemistry (Formalin-fixed paraffin-embedded sections)
Blocking step:
None
Antigen retrieval step:
Heat mediated
Incubation time:
30 minute(s)
Dilution: 1/800
Additional notes
The tissue was FFPE (formalin fixed, paraffin embedded tissue sectioned at 3-4 microns). The antigen retreival method was a heat-induced antigen retreival (HEIR) using a vegetable steamer. Sections were incubated in the steamer for 20 minutes in 1 mM EDTA buffer, pH 8.0. Sections were then rinsed in H2O and TBS buffer prior to starting the staining protocol.
The staining appeared to be very specific and robust, with very little to no non-specific background staining.
Staining Summary & Notes:
1mM EDTA Buffer, pH 8.0, HIER for 20 minutes in steamer
Rinse with TBS containing 0.05% Tween 20
10 minute blocking in 3% Peroxide
Rinse with TBS containing 0.05% Tween 20
30 minutes in Primary Antibody @ RT
Rinse with TBS containing 0.05% Tween 20
30 minutes in Secondary Antibody @ RT
Rinse with TBS containing 0.05% Tween 20
10 minutes in DAB+ (DakoCytomation) @ RT
Water Rinse
Counterstain for 6 minutes in Mayer's Hematoxylin (DakoCytomation) @ RT
Secondary antibody
Name:
DakoCytomation Envision+ Anti-Mouse/HRP (RTU)
Conjugation: HRP
Dilution: 1/1
