The sections were incubated with rabbit polyclonal antibody against MEnk (AB5026 from Millipore, St. Louis, MO, USA; 1:100, 1:1,000, or 1:10,000) or without the anti-MEnk antibody for 18 h in PBS-BSA. After several rinses in PBS, they were incubated with the polymer staining reagent by using the Histofine Simple Stain Kit (Nichirei, Tokyo, Japan) for 30 min. After several rinses in PBS, the bound peroxidase was visualized by incubating the sections with a solution containing 0.05% 3,3′-diaminobenzidine (DAB; Merck, Darmstadt, Germany) and 0.01% H2O2 in 0.05 M Tris-HCl (pH 7.4) for 10 min. After several rinses in water, the immunostained sections were dehydrated and cover-slipped with Malinol (Muto Pure Chemicals, Tokyo, Japan).
Ab5026
Manufactured by Merck Group
Sourced in United States
AB5026 is a laboratory equipment product manufactured by Merck Group. It is a specialized device designed for conducting precise scientific experiments and analyses. The core function of AB5026 is to provide accurate and reliable measurements and data collection for research and development purposes.
Automatically generated - may contain errors
Lab products found in correlation
Malinol, by Muto Pure Chemicals (3 mentions)
Histofine simple stain kit, by Nichirei Biosciences (2 mentions)
Vectastain elite abc reagent, by Vector Laboratories (2 mentions)
Dab 3 3 diaminobenzidine, by Merck Group (1 mentions)
3 3 diaminobenzidine (dab), by Merck Group (1 mentions)
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
Biotinyl tyramide, by PerkinElmer (1 mentions)
Biotinyl tyramide stock solution, by PerkinElmer (1 mentions)
Plus amplification diluent, by PerkinElmer (1 mentions)
Tsa biotin system, by PerkinElmer (1 mentions)
Mab354, by Merck Group (1 mentions)
A11012, by Thermo Fisher Scientific (1 mentions)
Ab22560, by Abcam (1 mentions)
A21448, by Thermo Fisher Scientific (1 mentions)
4 protocols using ab5026
1
Immunohistochemical Localization of MEnk
2
Immunohistochemical Localization of MEnk
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The sections were incubated with rabbit polyclonal antibody against MEnk (AB5026 from Millipore; 1:200, 1:2,000, or 1:20,000) or without the anti-MEnk antibody for 18 h in PBS-BSA. After several rinses in PBS, the sections were incubated with secondary biotinylated antibodies against rabbit IgG (Vector; 1:200) for 30 min. After several rinses in PBS, they were incubated with the Vectastain Elite ABC reagent for 30 min (Vectastain Elite ABC Kit; Vector). After several rinses in PBS, the bound peroxidase was visualized by incubating the sections with a solution containing 0.05% DAB (Merck) and 0.01% H2O2 in 0.05 M Tris-HCl (pH 7.4) for 10 min. After several rinses in water, the immunostained sections were dehydrated and cover-slipped with Malinol (Muto Pure Chemicals).
3
Immunostaining Protocol for Methionine-Enkephalin
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The sections were incubated with rabbit polyclonal antibody against MEnk (AB5026 from Millipore; 1:50,000, 1:500,000, or 1:5,000,000) or without the anti-MEnk antibody for 18 h in PBS-BSA. After several rinses in PBS, the sections were incubated with the polymer staining reagent (Histofine Simple Stain Kit; Nichirei, Tokyo, Japan) for 30 min. After several rinses in PBS, they were processed for tyramide signal amplification (TSA) using the TSA Biotin System (Perkin Elmer, Boston, MA, USA). The sections were incubated in Biotinyl Tyramide (amplification reagent) Working Solution that was made by diluting Biotinyl Tyramide Stock Solution (Perkin Elmer) 1:50 using 1X Plus Amplification Diluent (Perkin Elmer, FP1135) for 30 min. After several rinses in PBS, the sections were incubated with the Vectastain Elite ABC reagent (Vector) for 30 min in PBS. After several rinses in PBS, the bound peroxidase was visualized by incubating the sections with a solution containing 0.05% DAB and 0.01% H2O2 in 0.05M Tris-HCl (pH 7.4) for 10 min. After several rinses in water, the immunostained sections were dehydrated and cover-slipped with Malinol (Muto Pure Chemicals). Overview protocol for the PBTA-DAB staining is shown in Figure 1 (left).
4
Immunodetection of Neuropeptides in Tissue
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Calcitonin gene-related peptide (CGRP), somatostatin (SST) and methionine-enkephalin (met-enk) immunoreactivities were respectively detected using sheep polyclonal anti-CGRP (Abcam Ab22560, 1:2000) , rat monoclonal anti-SST (Millipore MAB354, 1:1000) and rabbit polyclonal anti-met-enk (Millipore AB5026, 1:1000) antibodies. Secondary antibodies were AlexaFluor 647-conjugated donkey anti-sheep (Molecular Probes A-21448, 1:500) for CGRP, Dylight 650-conjugated goat anti-rat (Thermofisher SA5-10021, 1:500) for SST, AlexaFluor 488-conjugated goat anti rabbit (Molecular Probes A-11034, 1:2000) for metenk. For colocalization studies between mu-Cherry and CGRP or SST, the fluorescent signal associated with the mu-mCherry fusion construct was amplified using rabbit polyclonal antidsred (Clontech 632496, 1:1000) and AlexaFluor 594-conjugated goat anti rabbit (Molecular Probes A-11012, 1:2000) antibodies. The absence of cross-reactivity (rabbit/sheep, rabbit/rat) was checked in control experiments. Immunohistochemistry was also performed without primary antibodies to verify absence of non-specific staining by the secondary antibody alone.
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