Timed embryos and postnatal pups were collected at designated day. The head samples were dissected in ice-cold phosphate buffer saline (PBS) and fixed in 4% paraformaldehyde (PFA)/PBS overnight at 4°C. Samples were then dehydrated in gradient sucrose/PBS washes and embedded in OCT. Frozen samples were sectioned in 8 μm. Alkaline phosphatase-Alcian blue (AP-AB) staining was performed as described previously (Dong et al., 2019 (link)). In short, sections were incubated with 0.03% nitro-blue tetrazolium chloride (NBT) (Roche, 11383213001) and 0.02% 5-bromo-4-chloro-3-indolyphosphate p-toluidine salt (BCIP) (Roche, 11383221001) in NTMT solution (100 mM NaCl, 100 mM Tris pH 9.5, 50 mM MgCl2 and 0.1% Tween-20 in distilled water). When AP signal was fully developed, slides were rinsed with distilled water then were immersed in 1% alcian blue 8GX (Sigma, A5268) in 0.1 N HCl, and then counterstained with 0.1% nuclear fast red (Acros Organics, 211980050) in 0.08 M aluminum sulfate (Al2(SO4)3·18H2O).
5 bromo 4 chloro 3 indolyphosphate p toluidine salt bcip
Manufactured by Roche
Sourced in Japan
5-bromo-4-chloro-3-indolyphosphate p-toluidine salt (BCIP) is a colorimetric substrate used for the detection of alkaline phosphatase activity in various biochemical and molecular biology applications.
Automatically generated - may contain errors
Lab products found in correlation
Nitro blue tetrazolium chloride, by Roche (1 mentions)
Nuclear fast red, by Thermo Fisher Scientific (1 mentions)
Alcian blue 8gx, by Merck Group (1 mentions)
Bovine serum, by Thermo Fisher Scientific (1 mentions)
Rabbit serum, by Thermo Fisher Scientific (1 mentions)
Alkaline phosphatase labeling kit nh2, by Dojindo Laboratories (1 mentions)
Nitrocellulose membrane ff120 hp, by GE Healthcare (1 mentions)
2 morpholinoethanesulfonicacid mes, by Dojindo Laboratories (1 mentions)
1 ethyl 3 3 dimethylaminopropyl carbodiimide hydrochloride, by GE Healthcare (1 mentions)
N hydroxysuccinimide, by GE Healthcare (1 mentions)
Colloidal gold, by BBI Solutions (1 mentions)
Tris hydroxymethyl aminomethane tris, by Nacalai Tesque (1 mentions)
2 protocols using 5 bromo 4 chloro 3 indolyphosphate p toluidine salt bcip
1
Alkaline Phosphatase-Alcian Blue Staining
2
Immunoassay Reagents and Materials Protocol
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The reagents used in this study were as follows: Nitrocellulose membrane FF120 HP, glass filter, 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC), and N-hydroxysuccinimide (NHS) purchased from GE Healthcare (IL, USA); casein and NaN3 purchased from Kanto Chemicals (Tokyo, Japan); Tris (hydroxymethyl) aminomethane (Tris) and N-cyclohexyl-3-aminopropanesulfonic acid (CAPS) purchased from Nacalai Tesque (Kyoto, Japan); and absorption pad and Bovine serum albumin (BSA) bought from Merck (NJ, USA). The backing sheet was obtained from Nippn Engineering (Tokyo, Japan). Colloidal gold was purchased from BBI Solutions, (Newport, UK). A microtiter plate (96 well) was obtained from Greiner (Frickenhausen, Germany). Alkaline Phosphatase Labeling Kit-NH2, 3-[(3-Cholamidopropyl) dimethylammonio]-propanesulfonate (CHAPS), and 2-morpholinoethanesulfonic acid (MES) were purchased from Dojindo (Kumamoto, Japan).
5-Bromo-4-chloro-3-indolyphosphate p-toluidine salt (BCIP) was purchased from Roche Diagnostics (Basel, Switzerland). Colored cellulose particles (NanoAct TM ) were obtained from Asahi Kasei (Tokyo, Japan). Bovine serum and rabbit serum were purchased from Thermo Fisher Scientific (Tokyo, Japan). Other reagents were obtained from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan).
5-Bromo-4-chloro-3-indolyphosphate p-toluidine salt (BCIP) was purchased from Roche Diagnostics (Basel, Switzerland). Colored cellulose particles (NanoAct TM ) were obtained from Asahi Kasei (Tokyo, Japan). Bovine serum and rabbit serum were purchased from Thermo Fisher Scientific (Tokyo, Japan). Other reagents were obtained from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan).
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