Adult worm pairs were fixed in Bouin's solution (picric acid/acetic acid/formaldehyde; 15/1/5) and embedded in paraplast (Paraplast plus, Sigma). Sections (5 μm) were incubated in xylol to remove paraplast. Following rehydration, slides were treated with proteinase K (1 μg/ml) and sections dehydrated. For hybridization, in vitro transcripts (corresponding to nt 616–1082 sequence of SmShb (Genbank Accession Number JN864885)) were synthesized and labeled with digoxigenin following the protocol of the manufacturer (Roche). The correct size of labeled sense and antisense transcripts was controlled by gel electrophoresis and the quality of RNA probes was checked by blotting and detection of digoxigenin using alkaline phosphatase-conjugated anti-digoxigenin antibodies, naphtol-AS-phosphatase, and Fast Red TR (Sigma). In situ hybridization was performed at 57°C for 16 h as previously described [24 (link)]. Sections were washed in 0.5×SSC (75 mM NaCl, 7.5 mM sodium citrate, pH 7) and detection of alkaline phosphatase on slides was achieved as described above.
Fast red tr
Manufactured by Merck Group
Sourced in Germany
Fast Red TR is a laboratory reagent used in various applications. It serves as a chromogenic substrate that undergoes a color change reaction when exposed to specific enzymes, allowing for the detection and visualization of target analytes. The product's core function is to provide a colorimetric signal for analytical purposes, but its specific intended use should not be extrapolated.
Automatically generated - may contain errors
Lab products found in correlation
Naphthol as mx phosphate, by Merck Group (10 mentions)
Paraplast plus, by Merck Group (3 mentions)
Napthol as tr phosphate solution, by Merck Group (2 mentions)
Napthol as mx phosphate, by Merck Group (2 mentions)
Naphthol as tr phosphate, by Merck Group (2 mentions)
Naphtol astr phosphate, by Merck Group (2 mentions)
Naphtol as phosphatase, by Merck Group (2 mentions)
Tris maleate buffer, by Merck Group (2 mentions)
Tris buffer, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
N n dimethylformamide (dmf), by Merck Group (1 mentions)
Shandon cryomatrix, by Thermo Fisher Scientific (1 mentions)
Rhbmp 2, by R&D Systems (1 mentions)
Rhbmp 2 enzyme linked immunosorbent assay elisa kit, by R&D Systems (1 mentions)
Cell counting kit 8 cck 8, by Merck Group (1 mentions)
Mem non essential amino acid, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
L glutamine, by Merck Group (1 mentions)
Foetal bovine serum (fbs), by Merck Group (1 mentions)
31 protocols using fast red tr
1
In Situ Hybridization of Schistosoma Shb Gene
2
Cellular Staining with Fast Red TR
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Cells were washed with phosphate buffer saline, fixed with 10% neutral buffered formalin, and stained for 30 min using 120 mM Tris buffer (Sigma-Aldrich) containing 1.8 mM fast red TR (Sigma-Aldrich) and 0.9 mM naphthol AS-MX phosphate (Sigma-Aldrich) at 37°C.
3
Osteogenic Lineage Induction and Staining
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Cells were induced to osteogenic lineage and fixed with acetone/citrate buffer pH 4.2 (1.5:1) for 5 min at room temperature. Cells were stained with Napthol-AS-TR-phosphate solution (Sigma-Aldrich ApS) for 1 h at room temperature. Napthol-AS-TR-phosphate solution consists of Napthol-AS-TR-phosphate diluted 1:5 in H2O and Fast Red TR (Sigma-Aldrich ApS) diluted 1:1.2 in 0.1 M Tris buffer, pH 9.0, after which both solutions were mixed 1:1 and applied to the cells.
4
Alkaline Phosphatase Activity Assay
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Cells were incubated with naphthol AS-TR phosphate solution containing Fast Red TR (Sigma-Aldrich) as described previously [27 (link)]. Alkaline phosphatase activity was measured using p-nitrophenyl phosphate (Fluka Chemie) as substrate [28 (link)].
5
Fluorescent Staining for Cell Analysis
6
Quantifying Bone Morphogenetic Protein in CHO Cells
7
Alkaline Phosphatase Activity Assay
8
Synthesis and Characterization of Functionalized Nanoparticles
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2-Methyl-2-oxazoline, NaOH pellets, phosphate buffer saline (PBS) tablets, foetal bovine serum (FBS), 2-mercaptosuccinic acid (MSA) (97%), sodium borohydride (NaBH4), nitric acid (70%) were purchased from Sigma-Aldrich Australia and used as received. Hydrochloric acid (36%, Ajax Finechem Pty. Ltd. Australia) was used as received. Dulbecco's Modified Eagle Medium (DMEM) was purchased from Gibco. Penicillin, streptomycin, l -glutamine, MEM non-essential amino acids, cell counting kit-8 (CCK-8), Naphthol AS-MX phosphate and Fast Red TR were obtained from Sigma. Silver nitrate (AgNO3) and glass coverslips were bought from ProSciTech, Australia. Ultra-pure MilliQ water (resistivity 18.2 Ω) was used for all experiments and cleaning procedures.
9
Assaying Acid Phosphatase Activity in Roots
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Seeds were germinated in vertical gels so that roots could grow on the gel surface. To determine the total activity of acid phosphatases, roots from each seedling were submerged in 2ml of assay medium for 15min and activity was determined by measuring the absorbance at 550nm. The assay medium contained 50mM trisodium citrate buffer (pH 5.6), 2.5mM Fast Red TR (Sigma-Aldrich), and 3.4mM 1-naphthyl phosphate, which is an artificial substrate (Dinkelaker and Marschner, 1992 ).
10
In Situ Hybridization for Fhabl1 Transcripts
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To detect the occurrence of transcripts of Fhabl1, in situ hybridization was performed as described earlier with slight modifications (31 (link)). Twelve-week-old F. hepatica worms were embedded in Tissue-Tek (Sakura Finetek), frozen on dry ice, and stored at −80°C until use. Traversal cryosections of 10 μm thickness were prepared using a cryostat HM525 (Thermo Fisher Scientific, Germany). Sections were dried, post-fixed in 4% PFA, and permeabilized with PBSTx. The hybridization reaction was carried out overnight at 55°C. Following washing with saline sodium citrate buffer (SSC), the sections were incubated with anti-DIG antibodies coupled with alkaline phosphatase (Roche, Germany). After subsequent washing steps with maleic acid buffer with Tween (MAB-T), the development reaction was carried out using naphthol-AS-phosphate, and Fast Red TR (Sigma). The Riboprobes for the hybridization reaction were generated as previously described (32 (link)). The following primers were used to generate the template for probe synthesis with a length of 596 bp: 5′-GAATCTCCTTCTCCTAACGGT-3′, reverse 5′-ACCAGATTTTTTAGGAGGTCTTC-3′. The labeling reaction using digoxigenin-11-UTP (NU-803-DIGXS; Jena Bioscience, Germany) was done using T3 or SP6 RNA polymerases for sense and antisense probes. Labeled transcripts were controlled for the correct size by gel electrophoresis.
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