Western blot analysis of p16 ARC on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (bsm-52302R, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: MCF-7 cell lysate Lane 2: SK-Br-3 cell lysate

Blocking buffer: 5% NFDM/TBST Primary Ab dilution: 1:2000 Primary Ab incubation condition: 2 hours at room temperature Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: 1: HeLa, 2: HCT-116, 3: SW480, 4: Rat brain, 5: Mouse brain, 6: Mouse ovary Protein loading quantity: 20 μg Exposure time: 60 s Predicted MW: 16 kDa Observed MW: 16 kDa

Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-p16 ARC antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (bsm-52302R, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-p16 ARC antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (bsm-52302R, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-p16 ARC antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (bsm-52302R, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-p16 ARC antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (bsm-52302R, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Tissue: Human endometrium carcinoma Section type: Formalin fixed & Paraffin - embedded section Retrieval method: High temperature and high pressure Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary Ab dilution: 1:100 Primary Ab incubation condition: 1 hour at room temperature Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to use) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for bsm-52302R

Tissue: Human spleen Section type: Formalin fixed & Paraffin - embedded section Retrieval method: High temperature and high pressure Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary Ab dilution: 1:100 Primary Ab incubation condition: 1 hour at room temperature Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to use) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for bsm-52302R

ICC staining of p16 ARC in N2A cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (bsm-52302R, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Cell line: Neuro-2a Fixative: 4% Paraformaldehyde Permeabilization: 0.1% TritonX-100 Primary Ab dilution: 1:50 Primary incubation condition: 4°C overnight Secondary Ab: Goat Anti-Rabbit IgG Nuclear counter stain: DAPI (Blue) Comment: Color green is the positive signal for bsm-52302R