[IF=5.11] He,et al.Disposable Morpho menelaus Based Flexible Microfluidic and Electronic Sensor for the Diagnosis of Neurodegenerative Disease.(2018) Advanced Healthcare Materials. 7:.  microfluidic chip ;  Human.  

PubMed:29345124

Sample: Brain (Mouse) Lysate at 40 ug
Primary: Anti- AD7c-NTP(bs-0046R) at 1/300 dilution
Secondary: HRP conjugated Goat-Anti-rabbit IgG (bs-0295G-HRP) at 1/5000 dilution
Predicted band size: 41 kD
Observed band size: 41 kD

Sample:
Lane 1: Mouse Cerebrum tissue lysates
Lane 2: Rat Cerebellum tissue lysates
Primary: Anti-Neuronal thread protein AD7c-NTP (bs-0046R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 41 kDa
Observed band size: 43 kDa

Paraformaldehyde-fixed, paraffin embedded (rat cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Neuronal thread protein AD7c-NTP) Polyclonal Antibody, Unconjugated (bs-0046R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Tissue/cell: human brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-AD7c-NTP Polyclonal Antibody, Unconjugated(bs-0046R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Tissue/cell: Mouse brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-AD7c-NTP Polyclonal Antibody, Unconjugated(bs-0046R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Blank control: SHSY5Y.
Primary Antibody (green line): Rabbit Anti-Neuronal thread protein AD7c-NTP antibody (bs-0046R)
Dilution: 1ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.