DEC-2005-IHC Envision+ method: Difference between revisions
Daniela Elena Costea 2005, Oral Pathology, Haukeland Hospital, Bergen, Norway. PROTOCOL: IHC EnVision+ method for E-cadherin monoclonal antibody (R&D) on frozen sections of normal human oral mucosa (NHOM) and organotypic cultures of NHOM ~ 4 h |
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[[Category:Protocols]] | |||
[[Category: Daniela Elena Costea]] | [[Category: Daniela Elena Costea]] | ||
[[Category: Oral Pathology Dept.]] | [[Category: Oral Pathology Dept.]] | ||
[[Category: Haukeland Hospital]] | [[Category: Haukeland Hospital]] | ||
[[Category: Norway]] | [[Category: Norway]] | ||
Latest revision as of 03:09, 30 August 2018
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Headline1|Headline2|Headline3
Aceton cold 50%|In ventilation|30 s
Aceton cold 100%|In ventilation |5 min
Wash|TBS|2 x 5 min
Primary antibody |E-cadherin in humidity chamber
1/9000 diluted in TBS |60 min
Wash|TBS + TWEEN 20
TBS: 500ml of the stock
(6.06 g TRIS/HCl from Merck L479219-136 + 1.39 g TRIS/base from Sigma
T1503 in 1ldH2O check pH7.6) in 5l NaCl 0.87% (87g NaCl from Merck K33224404-418 in 10 l dH2O)|2 x 5 min
Inactivation of peroxidase|peroxidase block from the EnVision+ kit|5 min
Wash|TBS|2 x 5 min
Secondary antibody|EnVision+ mouse
(DAKO)|30 min
Wash |TBS ± TWEEN 20|2 x 5 min
Visualisation|DAB
20 ul + 1 ml buffer
(1 drop DAB+ 1 ml buffer)|5 min
Wash|Running water|10 min
Counterstain|Hematoxylin|4 min
Wash|Running water |10 min
Alchool|70 %|30 s
Alchool|96 %|2 min
Alchool|Absolut|2 min
Toluen, xylol|In ventilation|5 min
Mounting of sections|Eukit|20 min
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