Digoxigenin-labeled RNA probes were transcribed with T7, T3 or SP6 polymerase (Ambion, Life Technologies, United States). WISH was performed as described previously (Thisse and Thisse, 2008 (link)). The probes labeled by digoxigenin were detected using alkaline phosphatase coupled anti-digoxigenin Fab fragment antibody (Roche) with 5-bromo-4-chloro-3-indolyl-phosphate nitro blue tetrazolium staining (Vector Laboratories, Burlingame, CA, United States).
5 bromo 4 chloro 3 indolyl phosphate nitro blue tetrazolium staining
Manufactured by Vector Laboratories
Sourced in United States
5-bromo-4-chloro-3-indolyl-phosphate nitro blue tetrazolium staining is a chromogenic substrate used for the detection of alkaline phosphatase enzyme activity in biological samples. It produces a blue-purple colored precipitate upon enzymatic cleavage.
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Lab products found in correlation
3 protocols using 5 bromo 4 chloro 3 indolyl phosphate nitro blue tetrazolium staining
1
Whole-Mount In Situ Hybridization Protocol
2
Whole-Mount In Situ Hybridization Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Digoxigenin-labeled RNA probes were transcribed with T7, T3 or SP6 polymerase (Ambion, Life Technologies, USA). WISH was performed as described previously [51 (link)]. The probes labeled by digoxigenin were detected using alkaline phosphatase coupled anti-digoxigenin Fab fragment antibody (Roche) with 5-bromo-4-chloro-3-indolyl-phosphate nitro blue tetrazolium staining (Vector Laboratories, Burlingame, CA, USA).
15–30 embryos were used for each probe. The positive signals were counted under a microscope, and the mean value was obtained from the counts of all of the embryos in the same group.
15–30 embryos were used for each probe. The positive signals were counted under a microscope, and the mean value was obtained from the counts of all of the embryos in the same group.
3
Whole-Mount In Situ Hybridization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Digoxigenin-labeled RNA probes were transcribed with T7, T3, or SP6 polymerase (Ambion, Life Technologies, Carlsbad, CA). WISH was performed as described previously (Thisse and Thisse, 2008 (link)). The probes labeled by digoxigenin were detected using alkaline phosphatase coupled anti-digoxigenin Fab fragment antibody (Roche, Basel, Switzerland) with 5-bromo-4-chloro-3-indolyl-phosphate nitro blue tetrazolium staining (Vector Laboratories, Burlingame, CA). Ten to 30 embryos were used for each probe. The positive signals were counted under a microscope, and the mean value was obtained from the counts of all of the embryos in the same group.
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