Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Bone and cartilage)
Antigen retrieval step
Other
Permeabilization
No
Specification
Bone and cartilage
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative
70 % Ethanol
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1% BSA in PBS
Dilution
1/1000
Secondary antibody
Dilution
1/300
Name
Non-Abcam antibody was used: Polyclonal Goat Anti-Mouse Immunoglobulins/Biotiny
Host species: Goat
Clonality: Polyclonal
Conjugation: Biotin
Host species: Goat
Clonality: Polyclonal
Conjugation: Biotin
Additional data
Additional Notes
Tissue fixed in 70 % ethanol, embedded in MMA (not paraffin-embedded), cut in 7um sections
Rinsed in dH2O
Acetic acid 1 % - 10 min
Rinsed in dH2O
3 % hydrogen peroxide - 20 min
Rinsed in dH2O
10% BSA for 20min at RT.
Primary Antibody diluted 1/1000 in 1%BSA/PBS and incubated overnight at 4oC
Wash 3x in PBS – 10 min
Secondary Antibody - Biot. Goat-antimouse (Dako E433) 1:300 – 60 min at RT.
Wash 3x in PBS – 10 min
StreptAvidin (Dako P0397) 1:300 – 60 min
Rinsed in PBS and dH2O
AEC – in dark nr. A6926 - 30 min
Rinsed in dH2O
Counterstained with Mayers HE-staining for 10-30 sec. at RT.
Rinsed in dH2O
Mounted.
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投稿 Aug 26 2014