HA601131

p16 Recombinant Mouse Monoclonal Antibody [PD01-16]

cat.: HA601131

Product Type:	Recombinant Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human
Applications:	IHC-P, WB, IF-Cell
Clonality:	Monoclonal
Clone number:	PD01-16

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 16 kDa
Isotype:	IgG1
Immunogen:	Synthetic peptide.
Positive control:	Human tonsil tissue, human cervix poorly differentiated adenocarcinoma tissue, human cervix highly differentiated squamous cell carcinoma tissue, human ovary Advanced serous carcinoma tissue, HeLa cell lysate, HepG2 cell lysate, HEK-293 cell lysate, NIH:OVCAR-3 cell lysate, HeLa.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: IHC-P WB IF-Cell	1:200-1:500 1:1,000 1:100
Uniprot #:	SwissProt: P42771 Human
Alternative names:	ARF CDK4 inhibitor p16 INK4 CDK4I CDKN2 CDKN2A Cell cycle negative regulator beta CMM2 Cyclin dependent kinase 4 inhibitor A Cyclin dependent kinase inhibitor 2A INK4 INK4a Melanoma p16 inhibits CDK4 MLM MTS 1 MTS1 Multiple tumor suppressor 1 p16 p16 INK4a p16INK4a p19 P19ARF TP16

Images

	Fig1: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-p16 antibody (HA601131) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA601131) at 1/500 dilution for 1 hour 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.
	Fig2: Immunohistochemical analysis of paraffin-embedded human cervix poorly differentiated adenocarcinoma tissue with Mouse anti-p16 antibody (HA601131) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA601131) at 1/500 dilution for 1 hour 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.
	Fig3: Immunohistochemical analysis of paraffin-embedded human cervix highly differentiated squamous cell carcinoma tissue with Mouse anti-p16 antibody (HA601131) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA601131) at 1/200 dilution for 1 hour 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.

Fig4: Immunohistochemical analysis of paraffin-embedded human ovary
Advanced serous carcinoma tissue with Mouse anti-p16 antibody (HA601131) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA601131) at 1/500 dilution for 1 hour 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.

Fig5: Western blot analysis of p16 on different lysates with Mouse anti-p16 antibody (HA601131) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HepG2 cell lysate
Lane 3: HEK-293 cell lysate
Lane 4: NIH:OVCAR-3 cell lysate

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 2 minutes;

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 (HA601131) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

Fig6: Immunocytochemistry analysis of HeLa cells labeling p16 with Mouse anti-p16 antibody (HA601131) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-p16 antibody (HA601131) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

Fig7: Immunocytochemistry analysis of HeLa cells labeling p16 with Mouse anti-p16 antibody (HA601131) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-p16 antibody (HA601131) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) 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.

Ki67 (HA721115, green) was stained at 1/250 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.