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p16INK4a Recombinant Mouse Monoclonal Antibody [PD01-16]

Usd: 350

Specification

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Catalog# HA601131

p16INK4a Recombinant Mouse Monoclonal Antibody [PD01-16]

Application

  • IHC-P

  • WB

  • IF-Cell

  • IF-Tissue

Reactivity

  • Human

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

p16INK4a Recombinant Mouse Monoclonal Antibody [PD01-16]

Antibody Type

Recombinant Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within N terminal Human p16INK4a.

Species Reactivity

Human

Validated Applications

IHC-P, WB, IF-Cell, IF-Tissue

Molecular Weight

Predicted band size: 16 kDa

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.

Conjugation

unconjugated

Clone Number

PD01-16

RRID

AB_3071853

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • IHC-P

  • 1:200-1:500

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

  • IF-Tissue

  • 1:500

Target

Function

This gene generates several transcript variants which differ in their first exons. At least three alternatively spliced variants encoding distinct proteins have been reported, two of which encode structurally related isoforms known to function as inhibitors of CDK4 kinase. The remaining transcript includes an alternate first exon located 20 Kb upstream of the remainder of the gene; this transcript contains an alternate open reading frame (ARF) that specifies a protein which is structurally unrelated to the products of the other variants. This ARF product functions as a stabilizer of the tumor suppressor protein p53 as it can interact with, and sequester, the E3 ubiquitin-protein ligase MDM2, a protein responsible for the degradation of p53. In spite of the structural and functional differences, the CDK inhibitor isoforms and the ARF product encoded by this gene, through the regulatory roles of CDK4 and p53 in cell cycle G1 progression, share a common functionality in cell cycle G1 control. This gene is frequently mutated or deleted in a wide variety of tumors, and is known to be an important tumor suppressor gene.

Background References

1. Okamoto A. et. al. Mutations and altered expression of p16INK4 in human cancer. Proc. Natl. Acad. Sci. U.S.A. 91:11045-11049(1994).

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P42771 Human

Synonyms

ARF antibody

CDK4 inhibitor p16 INK4 antibody

CDK4I antibody

CDKN2 antibody

CDKN2A antibody

Cell cycle negative regulator beta antibody

CMM2 antibody

Cyclin dependent kinase 4 inhibitor A antibody

Cyclin dependent kinase inhibitor 2A antibody

INK4 antibody

Expand

Images

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-p16INK4a antibody (<a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>) 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.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-p16INK4a antibody (HA601131) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Cervix poorly differentiated adenocarcinoma <br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>, 1/500, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1120" style="font-weight: bold;text-decoration: underline;">HA1120</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Cervix poorly differentiated adenocarcinoma
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA601131, 1/500, 1 hour at room temperature.
    Secondary antibody: HA1120, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Cervix well-differentiated squamous cell carcinoma <br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>, 1/200, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1120" style="font-weight: bold;text-decoration: underline;">HA1120</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Cervix well-differentiated squamous cell carcinoma
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA601131, 1/200, 1 hour at room temperature.
    Secondary antibody: HA1120, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: High-grade serous ovarian carcinoma<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>, 1/500, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1120" style="font-weight: bold;text-decoration: underline;">HA1120</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: High-grade serous ovarian carcinoma
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA601131, 1/500, 1 hour at room temperature.
    Secondary antibody: HA1120, 20 minutes at room temperature.

  • Western blot analysis of p16INK4a on different lysates with Mouse anti-p16INK4a antibody (<a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HepG2 cell lysate<br />Lane 3: HEK-293 cell lysate<br />Lane 4: NIH:OVCAR-3 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 16 kDa<br />Observed band size: 16 kDa<br /><br />Exposure time: 2 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of p16INK4a on different lysates with Mouse anti-p16INK4a 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.

  • Immunocytochemistry analysis of HeLa cells labeling p16INK4a with Mouse anti-p16INK4a antibody (<a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>) at 1/100 dilution.<br /><br />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-p16INK4a antibody (<a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) 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.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling p16INK4a with Mouse anti-p16INK4a 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-p16INK4a 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.

  • Immunocytochemistry analysis of HeLa cells labeling p16INK4a with Mouse anti-p16INK4a antibody (<a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>) at 1/100 dilution.<br /><br />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-p16INK4a antibody (<a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) 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.<br /><br />Ki67 (<a href="/products/HA721115" style="font-weight: bold;text-decoration: underline;">HA721115</a>, green) was stained at 1/250 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling p16INK4a with Mouse anti-p16INK4a 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-p16INK4a 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.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: High-grade serous ovarian carcinoma <br />Sample: Paraffin-embedded section<br /><br />Primary antibody dilution: 1/200<br /><br />Antigen retrieval: ER2<br />Platform: Leica Biosystems BOND&reg; RX

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: High-grade serous ovarian carcinoma
    Sample: Paraffin-embedded section

    Primary antibody dilution: 1/200

    Antigen retrieval: ER2
    Platform: Leica Biosystems BOND® RX

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Cervix well-differentiated squamous cell carcinoma <br />Sample: Paraffin-embedded section<br /><br />Primary antibody dilution: 1/200<br /><br />Antigen retrieval: ER2<br />Platform: Leica Biosystems BOND&reg; RX

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Cervix well-differentiated squamous cell carcinoma
    Sample: Paraffin-embedded section

    Primary antibody dilution: 1/200

    Antigen retrieval: ER2
    Platform: Leica Biosystems BOND® RX

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Ovary Advanced serous carcinoma <br />Sample: Paraffin-embedded section<br /><br />Primary antibody dilution: 1/200<br /><br />Antigen retrieval: HIER (CC1)<br />Platform: Ventana&reg; BenchMark ULTRA

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Ovary Advanced serous carcinoma
    Sample: Paraffin-embedded section

    Primary antibody dilution: 1/200

    Antigen retrieval: HIER (CC1)
    Platform: Ventana® BenchMark ULTRA

  • Application: Immunofluorescence (IF-tissue)<br /><br />Species: Human<br />Tissue: Tonsil<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× PBS<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA601131" style="font-weight: bold;text-decoration: underline;">HA601131</a>, 1/500, overnight at 4℃.<br />Secondary antibody: Goat Anti-Mouse IgG (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>), 1.5 hours at room temperature.

    Application: Immunofluorescence (IF-tissue)

    Species: Human
    Tissue: Tonsil
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× PBS
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA601131, 1/500, overnight at 4℃.
    Secondary antibody: Goat Anti-Mouse IgG (iFluor™ 488, HA1125), 1.5 hours at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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