Blocking Assay:Article Title: Effects of Hyperbaric Oxygen Therapy on Aquaporin Expression in the Salivary Glands : An In Vivo Study
Article Snippet: .. Then, the slides were immersed in a peroxidase blocking solution(BLOXALL® Endogenous Peroxidase and Alkaline Phosphatase Blocking Solution;Vector Laboratories, Newark, CA, USA)for 10 min, washed with PBS, and re‒ immersed in a blocking solution(2.5% normal horse serum;Vector Laboratories,)for 30 min. .. The sections were incubated overnight at 4°C with the primary antibody(Aquaporin‒1:rabbit anti‒Aquaporin‒1 and anti‒ Aquaporin‒5 polyclonal antibody 20333‒1‒AP;Proteintech, Rosemont, Aquaporin‒5:1:1,000, rabbit anti Aquaporin‒5 Polyclonal antibody BJ02243284;Bioss), which was diluted to 1:100 in 2.5% normal horse serum.
Article Title: Mouse skeletal muscle satellite cells co-opt the tenogenic gene Scleraxis to instruct regeneration
Article Snippet: Muscle sections were hydrated with PBS, permeabilized with 0.5% Triton X-100 (Sigma-Aldrich)/PBS (0.5% PBT) for 15 min, washed with 0.05% PBT, and blocked with MOM block (Vector Lab) overnight. .. Sections were washed and incubated in blocking solution (1 X carbo-free blocking solution (Vector Lab) and 10% goat serum in 0.05% PBT) for 2 hr at room temperature, followed by incubation with primary antibodies diluted in blocking solution overnight at 4 °C. ..
Article Title: Descending locus coeruleus noradrenergic signaling to spinal astrocyte subset is required for stress-induced mechanical pain hypersensitivity
Article Snippet: .. DOI: https://doi.org/10.7554/eLife.104453 16 of 24 spinal cord and brain sections (30 μm) were incubated in blocking solution (3% normal goat serum [#S- 1000; Vector Laboratories] or normal donkey serum [#017- 000- 121; Jackson ImmunoResearch]) for 2 hr at room temperature and then incubated for 48 hr at 4°C with primary antibodies: polyclonal rabbit anti- tyrosine hydroxylase (TH; 1:1000; #AB152; Millipore); polyclonal sheep anti- TH (1:1000; #AB1542; Millipore); monoclonal mouse anti- noradrenaline transporter (NET; 1:2000; #NET05- 2; Mab Technologies); polyclonal rabbit anti- green fluorescent protein (GFP; 1:1000; #598; MBL International); monoclonal rabbit anti- hemagglutinin (HA)- tag (1:1000; #3724; Cell Signaling); monoclonal rat antiglial fibrillary acidic protein (GFAP; 1:2000; #13- 0300; Invitrogen); polyclonal goat anti- SRY- related high- mobility group box 9 (SOX9; 1:1000; # AF3075; R&D Systems); polyclonal goat anti- paired box 2 (PAX2; 1:500; # AF3364; R&D Systems); monoclonal rat anti- mCherry (1:2000; #M11217; Thermo Fisher Scientific); polyclonal rabbit anti- phospho- p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (pERK; 1:500; #9101; Cell Signaling); anti- isolectin B4 (IB4)- biotin conjugate (1:1000; # I21414; Thermo Fisher Scientific); and monoclonal rabbit anti- c- FOS (1:5000; #226 008; Synaptic systems). .. After incubation, tissue sections were washed and incubated for 3 hr at room temperature with secondary antibodies (Alexa Fluor 488, 546 and/or 555; #A11001, A11008, A11035, A11056, A11081, S32351; Thermo Fisher Scientific: #ab150178; Abcam: DyLight 405; #705- 475- 147; Jackson ImmunoResearch).
Article Title: Descending locus coeruleus noradrenergic signaling to spinal astrocyte subset is required for stress-induced mechanical pain hypersensitivity
Article Snippet: After incubation, the tissues were embedded in OCT compound (#4583; Sakura Finetek Japan, Osaka, Japan) and stored at −25°C before use. .. Transverse spinal cord and brain sections (30 μm) were incubated in blocking solution (3% normal goat serum [#S-1000; Vector Laboratories] or normal donkey serum [#017-000-121; Jackson ImmunoResearch]) for 2 hr at room temperature and then incubated for 48 hr at 4°C with primary antibodies: polyclonal rabbit anti-tyrosine hydroxylase (TH; 1:1000; #AB152; Millipore); polyclonal sheep anti-TH (1:1000; #AB1542; Millipore); monoclonal mouse anti-noradrenaline transporter (NET; 1:2000; #NET05-2; Mab Technologies); polyclonal rabbit anti-green fluorescent protein (GFP; 1:1000; #598; MBL International); monoclonal rabbit anti-hemagglutinin (HA)-tag (1:1000; #3724; Cell Signaling); monoclonal rat anti-glial fibrillary acidic protein (GFAP; 1:2000; #13-0300; Invitrogen); polyclonal goat anti-SRY-related high-mobility group box 9 (SOX9; 1:1000; # AF3075; R&D Systems); polyclonal goat anti-paired box 2 (PAX2; 1:500; # AF3364; R&D Systems); monoclonal rat anti-mCherry (1:2000; # M11217 ; Thermo Fisher Scientific); polyclonal rabbit anti-phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (pERK; 1:500; #9101; Cell Signaling); anti-isolectin B4 (IB4)-biotin conjugate (1:1000; # I21414 ; Thermo Fisher Scientific); and monoclonal rabbit anti-c-FOS (1:5000; #226 008; Synaptic systems). .. After incubation, tissue sections were washed and incubated for 3 hr at room temperature with secondary antibodies (Alexa Fluor 488, 546 and/or 555; #A11001, A11008, A11035, A11056, A11081, S32351 ; Thermo Fisher Scientific: #ab150178; Abcam: DyLight 405; #705-475-147; Jackson ImmunoResearch).
Article Title: APP E590D mutation increases generation of Aβ and Aη peptides and exacerbates tauopathy
Article Snippet: .. Fixed cells were washed with PBS, incubated with blocking solution (0.2% Triton X-100, 3% normal goat serum) for 1 h, incubated with primary antibody at 4 °C overnight, washed three times with PBST, and incubated with Alexa-488 or Alexa-594-conjugated secondary IgG antibodies for 1 h at room temperature (Vector Laboratories, Burlingame, CA). .. The slides were then washed three times with PBST before being mounted with a fluorochrome mounting solution (Vector Laboratories).
Article Title: Mouse skeletal muscle satellite cells co-opt the tenogenic gene Scleraxis to instruct regeneration
Article Snippet: Muscle sections were hydrated with PBS, permeabilized with 0.5% Triton X- 100 (Sigma- Aldrich)/PBS (0.5% PBT) for 15 min, washed with 0.05% PBT, and blocked with MOM block (Vector Lab) overnight. .. Sections were washed and incubated in blocking solution (1 X carbo- free blocking solution (Vector Lab) and 10% goat serum in 0.05% PBT) for 2 hr at room temperature, followed by incubation with primary antibodies diluted in blocking solution overnight at 4 °C. ..
Incubation:Article Title: Mouse skeletal muscle satellite cells co-opt the tenogenic gene Scleraxis to instruct regeneration
Article Snippet: Muscle sections were hydrated with PBS, permeabilized with 0.5% Triton X-100 (Sigma-Aldrich)/PBS (0.5% PBT) for 15 min, washed with 0.05% PBT, and blocked with MOM block (Vector Lab) overnight. .. Sections were washed and incubated in blocking solution (1 X carbo-free blocking solution (Vector Lab) and 10% goat serum in 0.05% PBT) for 2 hr at room temperature, followed by incubation with primary antibodies diluted in blocking solution overnight at 4 °C. ..
Article Title: Descending locus coeruleus noradrenergic signaling to spinal astrocyte subset is required for stress-induced mechanical pain hypersensitivity
Article Snippet: .. DOI: https://doi.org/10.7554/eLife.104453 16 of 24 spinal cord and brain sections (30 μm) were incubated in blocking solution (3% normal goat serum [#S- 1000; Vector Laboratories] or normal donkey serum [#017- 000- 121; Jackson ImmunoResearch]) for 2 hr at room temperature and then incubated for 48 hr at 4°C with primary antibodies: polyclonal rabbit anti- tyrosine hydroxylase (TH; 1:1000; #AB152; Millipore); polyclonal sheep anti- TH (1:1000; #AB1542; Millipore); monoclonal mouse anti- noradrenaline transporter (NET; 1:2000; #NET05- 2; Mab Technologies); polyclonal rabbit anti- green fluorescent protein (GFP; 1:1000; #598; MBL International); monoclonal rabbit anti- hemagglutinin (HA)- tag (1:1000; #3724; Cell Signaling); monoclonal rat antiglial fibrillary acidic protein (GFAP; 1:2000; #13- 0300; Invitrogen); polyclonal goat anti- SRY- related high- mobility group box 9 (SOX9; 1:1000; # AF3075; R&D Systems); polyclonal goat anti- paired box 2 (PAX2; 1:500; # AF3364; R&D Systems); monoclonal rat anti- mCherry (1:2000; #M11217; Thermo Fisher Scientific); polyclonal rabbit anti- phospho- p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (pERK; 1:500; #9101; Cell Signaling); anti- isolectin B4 (IB4)- biotin conjugate (1:1000; # I21414; Thermo Fisher Scientific); and monoclonal rabbit anti- c- FOS (1:5000; #226 008; Synaptic systems). .. After incubation, tissue sections were washed and incubated for 3 hr at room temperature with secondary antibodies (Alexa Fluor 488, 546 and/or 555; #A11001, A11008, A11035, A11056, A11081, S32351; Thermo Fisher Scientific: #ab150178; Abcam: DyLight 405; #705- 475- 147; Jackson ImmunoResearch).
Article Title: Descending locus coeruleus noradrenergic signaling to spinal astrocyte subset is required for stress-induced mechanical pain hypersensitivity
Article Snippet: After incubation, the tissues were embedded in OCT compound (#4583; Sakura Finetek Japan, Osaka, Japan) and stored at −25°C before use. .. Transverse spinal cord and brain sections (30 μm) were incubated in blocking solution (3% normal goat serum [#S-1000; Vector Laboratories] or normal donkey serum [#017-000-121; Jackson ImmunoResearch]) for 2 hr at room temperature and then incubated for 48 hr at 4°C with primary antibodies: polyclonal rabbit anti-tyrosine hydroxylase (TH; 1:1000; #AB152; Millipore); polyclonal sheep anti-TH (1:1000; #AB1542; Millipore); monoclonal mouse anti-noradrenaline transporter (NET; 1:2000; #NET05-2; Mab Technologies); polyclonal rabbit anti-green fluorescent protein (GFP; 1:1000; #598; MBL International); monoclonal rabbit anti-hemagglutinin (HA)-tag (1:1000; #3724; Cell Signaling); monoclonal rat anti-glial fibrillary acidic protein (GFAP; 1:2000; #13-0300; Invitrogen); polyclonal goat anti-SRY-related high-mobility group box 9 (SOX9; 1:1000; # AF3075; R&D Systems); polyclonal goat anti-paired box 2 (PAX2; 1:500; # AF3364; R&D Systems); monoclonal rat anti-mCherry (1:2000; # M11217 ; Thermo Fisher Scientific); polyclonal rabbit anti-phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (pERK; 1:500; #9101; Cell Signaling); anti-isolectin B4 (IB4)-biotin conjugate (1:1000; # I21414 ; Thermo Fisher Scientific); and monoclonal rabbit anti-c-FOS (1:5000; #226 008; Synaptic systems). .. After incubation, tissue sections were washed and incubated for 3 hr at room temperature with secondary antibodies (Alexa Fluor 488, 546 and/or 555; #A11001, A11008, A11035, A11056, A11081, S32351 ; Thermo Fisher Scientific: #ab150178; Abcam: DyLight 405; #705-475-147; Jackson ImmunoResearch).
Article Title: APP E590D mutation increases generation of Aβ and Aη peptides and exacerbates tauopathy
Article Snippet: .. Fixed cells were washed with PBS, incubated with blocking solution (0.2% Triton X-100, 3% normal goat serum) for 1 h, incubated with primary antibody at 4 °C overnight, washed three times with PBST, and incubated with Alexa-488 or Alexa-594-conjugated secondary IgG antibodies for 1 h at room temperature (Vector Laboratories, Burlingame, CA). .. The slides were then washed three times with PBST before being mounted with a fluorochrome mounting solution (Vector Laboratories).
Article Title: Mouse skeletal muscle satellite cells co-opt the tenogenic gene Scleraxis to instruct regeneration
Article Snippet: Muscle sections were hydrated with PBS, permeabilized with 0.5% Triton X- 100 (Sigma- Aldrich)/PBS (0.5% PBT) for 15 min, washed with 0.05% PBT, and blocked with MOM block (Vector Lab) overnight. .. Sections were washed and incubated in blocking solution (1 X carbo- free blocking solution (Vector Lab) and 10% goat serum in 0.05% PBT) for 2 hr at room temperature, followed by incubation with primary antibodies diluted in blocking solution overnight at 4 °C. ..
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