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Blank control (blue): Hep G2 Cells (fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice). Primary Antibody: Rabbit Anti-TNFR1/FITC Conjugated antibody, Dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG/FITC (orange), used under the same conditions. |
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Paraformaldehyde-fixed, paraffin embedded (Rat uterus), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (P21) Polyclonal Antibody, Unconjugated (orb158074) at 1:400 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining. |
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Tissue/Cell: human colon carcinoma, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37C for 20 min, Incubation: Anti-P21 Polyclonal Antibody, Unconjugated (orb158074) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |
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Tissue/Cell: human hepatoma tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37C for 20 min, Incubation: Anti-P21 Polyclonal Antibody, Unconjugated (orb158074) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |
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Tissue/Cell: human lung carcinoma, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37C for 20 min, Incubation: Anti-P21 Polyclonal Antibody, Unconjugated (orb158074) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |
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Tissue/Cell: HUVEC cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37C for 20 min, Antibody incubation with (CDKN1A/P21) Polyclonal Antibody, Unconjugated (orb158074) 1:100, 90 minutes at 37C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868805) at 37C for 90 minutes, DAPI (blue) was used to stain the cell nuclei. |