The sorghum 4th internode stem samples were embedded in paraffin and the following RNA hybridization process was carried out as previously described (Wang et al., 2021 (link)). DNA template for RNA probes was amplified from sorghum cDNA using gene-specific primers (see Supplementary Table S6
Anti fluorescein ap
Manufactured by Merck Group
Anti-Fluorescein-AP is a laboratory reagent used in various immunoassay techniques. It functions as a detection antibody, specifically binding to the fluorescein molecule and enabling the visualization of target analytes through enzymatic color development.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using anti fluorescein ap
1
RNA Hybridization of Sorghum Internode
2
Whole-mount in situ Hybridization Protocol
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Whole-mount in situ hybridization was performed as previously described (deCarvalho et al., 2014 (link); Gamse et al., 2002 (link)). In brief, larvae and dissected brains were fixed in 4% paraformaldehyde (P6148, Sigma-Aldrich) in 1 X PBS (phosphate-buffered saline) at 4 °C overnight. To synthesize RNA probes, the following restriction enzymes and RNA polymerases were used: lratd2a (BamHI/T7), fos (NotI/SP6), slc5a7a (NotI/SP6), kctd12.1 (EcoRI/T7) (deCarvalho et al., 2013 (link); Hong et al., 2013 (link)). Probes were labeled with UTP-digoxigenin (11093274910, Roche) and samples incubated at 70 °C in hybridization solution containing 50% formamide. Hybridized probes were detected using alkaline phosphatase-conjugated antibodies (Anti-Digoxigenin-AP, #11093274910, and Anti-Fluorescein-AP, #11426338910, Sigma-Aldrich) and visualized by staining with 4-nitro blue tetrazolium (NBT, #11383213001, Roche), 5-bromo-4-chloro-3-indolyl-phosphate (BCIP, #11383221001, Roche) and 2-(4-Iodophenyl)–3-(4-nitrophenyl)–5-phenyltetrazolium Chloride (INT, #I00671G, Fisher Scientific).
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
Whole-mount in situ hybridization was performed as previously described (deCarvalho et al., 2014; Gamse et al., 2002) (link). In brief, larvae and dissected brains were fixed in 4% paraformaldehyde (P6148, Sigma-Aldrich) in 1X PBS (phosphate-buffered saline) at 4 °C overnight. To synthesize RNA probes, the following restriction enzymes and RNA polymerases were used: lratd2a
(BamHI/T7), fos (NotI/SP6), slc5a7a (NotI/SP6), kctd12.1 (EcoRI/T7) (deCarvalho et al., 2013; Hong et al., 2013) (link). Probes were labeled with UTP-digoxigenin (11093274910, Roche) and samples incubated at 70 °C in hybridization solution containing 50% formamide. Hybridized probes were detected using alkaline phosphatase-conjugated antibodies (Anti-Digoxigenin-AP, #11093274910, and Anti-Fluorescein-AP, #11426338910, Sigma-Aldrich) and visualized by staining with 4-nitro blue tetrazolium (NBT, #11383213001, Roche), 5-bromo-4-chloro-3-indolylphosphate (BCIP, #11383221001, Roche) and 2-(4-Iodophenyl)-3-(4-nitrophenyl)-5phenyltetrazolium Chloride (INT, #I00671G, Fisher Scientific).
(BamHI/T7), fos (NotI/SP6), slc5a7a (NotI/SP6), kctd12.1 (EcoRI/T7) (deCarvalho et al., 2013; Hong et al., 2013) (link). Probes were labeled with UTP-digoxigenin (11093274910, Roche) and samples incubated at 70 °C in hybridization solution containing 50% formamide. Hybridized probes were detected using alkaline phosphatase-conjugated antibodies (Anti-Digoxigenin-AP, #11093274910, and Anti-Fluorescein-AP, #11426338910, Sigma-Aldrich) and visualized by staining with 4-nitro blue tetrazolium (NBT, #11383213001, Roche), 5-bromo-4-chloro-3-indolylphosphate (BCIP, #11383221001, Roche) and 2-(4-Iodophenyl)-3-(4-nitrophenyl)-5phenyltetrazolium Chloride (INT, #I00671G, Fisher Scientific).
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