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Paraformaldehyde-fixed, paraffin embedded (Human breast cancer); 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 (phospho-ERK5 ( Ser731+Thr733)) Polyclonal Antibody, Unconjugated (SL5486R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.
Tissue/cell: human myometrium 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,SLC0005) at 37∩ for 20 min;
Incubation: Anti-phospho-ERK5(Ser731+Thr733) Polyclonal Antibody, Unconjugated(SL5486R) 1:200, overnight at 4∑C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Tissue/cell: human kidney tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,SLC0005) at 37∩ for 20 min;
Incubation: Anti-phospho-ERK5(Ser731+Thr733) Polyclonal Antibody, FITC conjugated (SL5486R-FITC) 1:200, used at 1:200 dilution for 40 minutes at 37∑C.
Tissue/cell: human lung carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,SLC0005) at 37∩ for 20 min;
Incubation: Anti-phospho-ERK5(Ser731+Thr733) Polyclonal Antibody, Unconjugated(SL5486R) 1:200, overnight at 4∑C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated (SL0295G-Cy3)used at 1:200 dilution for 40 minutes at 37∑C.
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