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