In situ hybridization studies were performed as previously described [48 (link)]. Briefly, digoxigenin-labeled antisense RNA probes were synthesized from a cDNA clone for Ptgs2 (clone ID: 30059181, Thermo Fisher Scientific) with T7 RNA polymerase (New England Biolabs, Ipswich, MA) and DIG RNA Labeling Mix (Roche, Indianapolis, IN) and purified with NucAway Spin Columns (Life Technologies, Carlsbad, CA). Deparaffinized and Protease K-treated sections were hybridized with RNA probes and incubated with anti-digoxigenin alkaline phosphatase (AP)-conjugated antigen binding fragments (Fab) (Roche). Specific signals were visualized with nitro blue tetrazolium chloride (NBT) and 5-Bromo-4-chloro-3-indolyl phosphate (BCIP) (Roche) and sections were counterstained with methyl green.
Anti digoxigenin alkaline phosphatase conjugated antigen binding fragments fab
Manufactured by Roche
Anti-digoxigenin alkaline phosphatase (AP)-conjugated antigen binding fragments (Fab) is a laboratory reagent. It consists of antibody fragments that are conjugated to the enzyme alkaline phosphatase. This reagent is used for the detection of digoxigenin-labeled biomolecules in various laboratory applications.
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2 protocols using anti digoxigenin alkaline phosphatase conjugated antigen binding fragments fab
1
In situ hybridization analysis of Ptgs2 expression
2
In Situ Hybridization for Ptgs2 Expression
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In situ hybridization studies were performed as previously described (Manieri et al., 2012 (link)). Briefly, digoxigenin-labeled antisense RNA probes were synthesized from a cDNA clone for Ptgs2 (clone ID: 30059181, Thermo Fisher Scientific) with T7 RNA polymerase (New England Biolabs, Ipswich, MA) and DIG RNA Labeling Mix (Roche, Indianapolis, IN) and purified with NucAway Spin Columns (Life Technologies, Carlsbad, CA). Deparaffinized and protease K-treated sections were hybridized with RNA probes and incubated with anti-digoxigenin alkaline phosphatase (AP)-conjugated antigen binding fragments (Fab) (Roche). Specific signals were visualized with nitro blue tetrazolium chloride (NBT) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP) (Roche) and sections were counterstained with methyl green.
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