Tg(fli1a:GAL4FF)ubs3; Tg(UAS:bmpr1aaWT_IRES_EGFP)/Tg(UAS:bmpr1aap.R438H_IRES_EGFP); Tg(7xTCF-Xla.Sia:NLS-mCherry)ia5 embryos were treated with PTU (Sigma) at 24hpf, anaesthetized with 3-aminobenzoic acid ethyl ester (Tricaine) (Sigma) and fixed in 4% PFA at 120dpf.
3 aminobenzoic acid ethyl ester tricaine
Manufactured by Merck Group
Sourced in Netherlands
3-aminobenzoic acid ethyl ester (Tricaine) is a white crystalline solid used as a laboratory reagent. It is a central nervous system depressant that can induce anesthesia in aquatic organisms.
Automatically generated - may contain errors
Lab products found in correlation
Hygromycin b, by Thermo Fisher Scientific (1 mentions)
Rifampicin, by Merck Group (1 mentions)
Recombinant human ifn γ protein, by R&D Systems (1 mentions)
Tmp269, by Selleck Chemicals (1 mentions)
Trichostatin a tsa, by Selleck Chemicals (1 mentions)
H89 dihydrochloride, by Merck Group (1 mentions)
Tmp195, by Selleck Chemicals (1 mentions)
Pneumatic picopump, by World Precision Instruments (1 mentions)
Borosilicate glass capillary needles, by Harvard Apparatus (1 mentions)
4 protocols using 3 aminobenzoic acid ethyl ester tricaine
1
Transgenic Zebrafish for BMP Signaling
2
Pharmacological Inhibition of Kinases and Histone Deacetylases
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H-89 dihydrochloride (PKA/PKB/AKT1 kinase inhibitor), 3-aminobenzoic acid ethyl ester (tricaine), and rifampicin were purchased from Sigma-Aldrich, Zwijndrecht, The Netherlands. H-89 analog 97i was synthesized by the Leiden Academic Center for Drug Research, Division of Medicinal Chemistry, Leiden University, Leiden, The Netherlands. Pan-HDAC inhibitor Trichostatin A (TSA) and class IIa HDAC inhibitors TMP195 and TMP269 were purchased from Selleckchem, Munich, Germany. Hygromycin B was acquired from Life Technologies-Invitrogen, Bleiswijk, The Netherlands. Recombinant human IFN-γ protein was acquired from R&D Systems, Wiesbaden, Germany.
3
Zebrafish Husbandry and Experimentation
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The husbandry of adult zebrafish lines and all zebrafish experiments described in this study was in accordance with guidelines from the local animal welfare committee (DEC) of the university (License number: protocol 14,198), in compliance with the international guidelines specified by the EU Animal Protection Directive 2010/63/EU, and was conducted according to standard protocols (www.zfin.org ). There was no adult zebrafish sacrificed in this study. All experiments were done with zebrafish larvae developed within 5 dpf, therefore prior to the free-feeding stage and did not fall under animal experimentation law according to the EU Animal Protection Directive 2010/63/EU. Zebrafish eggs and larvae were cultured and grown at 28.5°C in egg water (60 g/ml Instant Ocean sea salts). Zebrafish larvae were anesthetized with egg water containing 0.02% buffered 3-aminobenzoic acid ethyl ester (Tricaine, Sigma-Aldrich, Netherlands) for bacterial infection and imaging experiments.
The ABTL wild type zebrafish strain, tlr2sa19423 mutant (ENU-mutagenized) and the offspring of its wild type siblings or the following transgenic lines: Tg (mpeg1:EGFP)gl22, tlr2+/+ Tg (mpeg1:mCherry-F);TgBAC (mpx: EGFP) and tlr2−/− Tg (mpeg1:mCherry-F);TgBAC (mpx: EGFP) were used for this study (38 (link)).
The ABTL wild type zebrafish strain, tlr2sa19423 mutant (ENU-mutagenized) and the offspring of its wild type siblings or the following transgenic lines: Tg (mpeg1:EGFP)gl22, tlr2+/+ Tg (mpeg1:mCherry-F);TgBAC (mpx: EGFP) and tlr2−/− Tg (mpeg1:mCherry-F);TgBAC (mpx: EGFP) were used for this study (38 (link)).
4
Zebrafish Xenograft Melanoma Model
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Zebrafish and embryos were raised, staged and maintained according to standard procedures. The Institutional Committee for Animal Welfare of the Leiden University Medical Center (LUMC) approved this study. Tg(Fli1:GFP) zebrafish embryos were dechorionated at two days post-fertilisation (dpf). Single cell suspensions of melanoma cells were prepared in PBS and kept at 4°C before implantation. The cell suspension was loaded into borosilicate glass capillary needles (1 mm O.D. × 0.78 mm I.D.; Harvard Apparatus) and the injections were performed using a Pneumatic Picopump and a manipulator (World Precision Instruments, Stevenage, UK). Dechorionated embryos were anaesthetised with 0.003% 3-amino benzoic acid ethyl estertricaine, (Sigma)] and mounted on 10 cm Petridishes coated with 1% agarose. Approximately 200 cells were injected at the duct of Cuvier (DoC). Implanted zebrafish embryos were maintained at 33°C. Zebrafish in the Non-Silencing (NS) + SB431542 group were treated with 1μM SB-431542 added to the eggwater. All implantations were repeated at least three times with at least 30 embryos per group.
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