Cyclin D1 Immunohistochemistry Staining

Fine sectioning (5 μm) was done in formalin-fixed paraffin-embedded specimens on poly-L-lysine (Sigma, St. Louis, MO, USA) coated slides, followed by immunohistochemistry (IHC). The antigen retrieval for Cyclin D1 was performed in 10 mM tris-EDTA buffer (pH-9.0) at 900W for 15 min/360W for 5 min. The sections were consecutively washed in Tris buffer saline (TBS; 0.1M; pH: 7.4) and then incubated with primary antibody of Primary rabbit monoclonal of Cyclin D1 (monoclonal, M3635, Dako cytomation Glostrup, Denmark; 1:200 dilution) overnight followed by the use of polymer based secondary antibody (Envision System peroxidase kit, DAKO, Carpinteria, CA) for 1 hr at room temperature after washing with TBS. Previous studies from our laboratory have mentioned the complete methodological details^{9 (link),10 (link)}.

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Tripathi R., Rath G., Jawanjal P., Bharadwaj M, & Mehrotra R. (2019). ≤ Cyclin D1 protein affecting global women’s health by regulating HPV mediated adenocarcinoma of the uterine cervix. Scientific Reports, 9, 5019.

Publication 2019

poly-L-lysine M3635 Envision System peroxidase kit

Antibody Antigen Cyclin d1 Dilution Edta Embedded paraffin Formalin Immunohistochemistry Lysine Peroxidase Poly Polymer Rabbit Saline Tris buffer

Corresponding Organization :

Other organizations : National Institute of Cancer Prevention and Research, Safdarjang Hospital

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Variable analysis

independent variables

Antigen retrieval method: 10 mM tris-EDTA buffer (pH-9.0) at 900W for 15 min/360W for 5 min

dependent variables

Expression of Cyclin D1 protein

control variables

Formalin-fixed paraffin-embedded specimens
Poly-L-lysine (Sigma, St. Louis, MO, USA) coated slides
Primary rabbit monoclonal of Cyclin D1 (monoclonal, M3635, Dako cytomation Glostrup, Denmark; 1:200 dilution) used as primary antibody
Polymer based secondary antibody (Envision System peroxidase kit, DAKO, Carpinteria, CA) used for 1 hr at room temperature

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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