|Detection of Human p21/CIP1/CDKN1A by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line untreated (-) or treated (+) with 1 μM camptothecin (CPT) for 16 hours. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human p21/CIP1/CDKN1A Monoclonal Antibody (Catalog # MAB1047), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for p21/CIP1/CDKN1A at approximately 21 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
p21/CIP1/CDKN1A in Human Breast Cancer Tissue. p21/CIP1/CDKN1A was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Mouse Anti-Human p21/|
CIP1/CDKN1A Monoclonal Antibody (Catalog # MAB1047) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Intracellular Staining by Flow Cytometry
|Detection of p21/Cip1/CDKN1A in MCF‑7 Human Cell Line by Flow Cytometry. MCF‑7 human breast cancer cell line either treated with 1 μM camptothecin (CPT) for 16 hours (filled histogram) or untreated (open histogram) was stained with Mouse Anti-Human p21/CIP1/CDKN1A Monoclonal Antibody (Catalog # MAB1047), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.|
p21CIP1, also called CIP1 (CDK-interacting protein 1) and CDKN1A, is a 21 kDa Cyclin/Cyclin-dependent kinase (Cdk) inhibitor that blocks cell cycle progression from G1 to S phase in the cell cycle. Because p21 is a transcriptional target of the p53 tumor suppressor, p21 expression increases in cells that contain stabilized p53 due to genotoxic stress exposure.
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