p16INK4a Recombinant Rabbit Monoclonal Antibody [SU0702]
cat.: ET1608-62
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: SU0702
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 16 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human p16INK4a aa 107-156 / 156.
Positive control: 293 cell lysate, SiHa cell lysate, HepG2, Hela, PC-3M, human colon carcinoma tissue, human cervical carcinoma tissue, human ovary carcinoma tissue.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:500-1:1,000
1:100-1:500
1:100-1:500
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: P42771 Human
Alternative names: CCM2 CDK4 inhibitor p16 INK4 CDK4I CDKN2 CDKN2A Cell cycle negative regulator beta CMM2 Cyclin dependent kinase 4 inhibitor A Cyclin dependent kinase inhibitor 2A (melanoma p16 inhibits CDK4) Cyclin Dependent Kinase Inhibitor 2A Cyclin dependent kinase inhibitor 2A isoform 4 Cyclin dependent kinase inhibitor 2A isoforms 1/2/3 Cyclin dependent kinase inhibitor p16 INK4 INK4A MLM MTS1 Multiple tumor suppressor 1 p14 p16 P16INK4 p16INK4a p19 p19Arf TP16
Images
ET1608-62_1.jpg Fig1: Western blot analysis of p16INK4a 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 (ET1608-62, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: 293 cell lysate
Lane 2: SiHa cell lysate
ET1608-62_2.jpg Fig2: Western blot analysis of p16INK4a on different lysates with Rabbit anti-p16INK4a antibody (ET1608-62) at 1/1,000 dilution.

Lane 1: HeLa-si NT cell lysate
Lane 2: HeLa-si p16INK4a cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 16 kDa
Observed band size: 16 kDa

Exposure time: 20 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-62) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.
ET1608-62_3.jpg Fig3: ICC staining of p16INK4a in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-62, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-62_4.jpg Fig4: Immunocytochemistry analysis of Hela cells labeling p16INK4a with Rabbit anti-p16INK4a antibody (ET1608-62) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-p16INK4a antibody (ET1608-62) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
ET1608-62_5.jpg Fig5: ICC staining of p16INK4a in PC-3M cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-62, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-62_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-p16INK4a antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-62, 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.
ET1608-62_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue using anti-p16INK4a antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-62, 1/100) 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.
ET1608-62_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human ovary carcinoma tissue using anti-p16INK4a antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-62, 1/100) 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.
ET1608-62_9.jpg Fig9: Flow cytometric analysis of p16INK4a was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1608-62, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.