Immunohistochemical Analysis of Cell Cycle Regulators

5-μm sections of paraffin embedded, formalin fixed, lung were stained for p16, p21, and p53 by immunohistochemistry as previously described. [23 (link)] Briefly, the sections were incubated in PBS containing 5% normal goat serum and 1% bovine serum albumin (BSA), then incubated with a 1:100 dilution of mouse anti-human p16 (Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-human p21 (Santa Cruz Biotechnology) or mouse anti-human p53 antibody (Santa Cruz Biotechnology) overnight at 4°C. The sections were washed, incubated with horseradish peroxidase (HRP)-conjugated goat anti-mouse or anti-rabbit IgG (Santa Cruz Biotechnology). After washing, the bound peroxidase activity was detected using a diaminobenzidine (DAB) substrate kit (Vector Laboratories, Burlingame, CA).

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Disayabutr S., Kim E.K., Cha S.I., Green G., Naikawadi R.P., Jones K.D., Golden J.A., Schroeder A., Matthay M.A., Kukreja J., Erle D.J., Collard H.R, & Wolters P.J. (2016). miR-34 miRNAs Regulate Cellular Senescence in Type II Alveolar Epithelial Cells of Patients with Idiopathic Pulmonary Fibrosis. PLoS ONE, 11(6), e0158367.

Publication 2016

mouse anti-human p16 rabbit anti-human p21 horseradish peroxidase (HRP)-conjugated goat anti-mouse or anti-rabbit IgG diaminobenzidine (DAB) substrate kit

Anti antibody Anti igg Bovine serum albumin Dilution Embedded paraffin Formalin Goat Horseradish peroxidase Human Immunohistochemistry Lung Mouse Peroxidase Rabbit Serum Vector

Corresponding Organization : Kyungpook National University Hospital

Top 5 similar protocols

Variable analysis

dependent variables

Immunohistochemical staining intensity of p16, p21, and p53

control variables

5-μm sections of paraffin embedded, formalin fixed, lung
Incubation in PBS containing 5% normal goat serum and 1% bovine serum albumin (BSA)
Incubation with a 1:100 dilution of mouse anti-human p16, rabbit anti-human p21, or mouse anti-human p53 antibody overnight at 4°C
Washing and incubation with horseradish peroxidase (HRP)-conjugated goat anti-mouse or anti-rabbit IgG
Washing and detection of bound peroxidase activity using a diaminobenzidine (DAB) substrate kit

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