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p57 Kip2 Recombinant Rabbit Monoclonal Antibody [JE02-08]

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Specification

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Catalog# HA722137

p57 Kip2 Recombinant Rabbit Monoclonal Antibody [JE02-08]

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

p57 Kip2 Recombinant Rabbit Monoclonal Antibody [JE02-08]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human p57 Kip2 aa 31-80 / 316.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Target Molecular Weight

Predicted band size: 32 kDa

Positive Control

HeLa cell lysate, HeLa treated with 50nM dexamethasone for 16 hours cell lysate, SH-SY5Y cell lysate, K-562 cell lysate, 293T cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, HeLa cells treated with 50nM dexamethasone for 16 hours, human placenta tissue, human brain tissue, mouse brain tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

JE02-08

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IF-Cell

  • 1:250

  • IHC-P

  • 1:200

Target

Function

Cyclin-dependent kinase inhibitor 1C (p57, Kip2), also known as CDKN1C, is a protein which in humans is encoded by the CDKN1C imprinted gene. Cyclin-dependent kinase inhibitor 1C is a tight-binding inhibitor of several G1 cyclin/Cdk complexes and a negative regulator of cell proliferation. Mutations of CDKN1C are implicated in sporadic cancers and Beckwith-Wiedemann syndrome suggesting that it is a tumor suppressor candidate. CDKN1C is a tumor suppressor human gene on chromosome 11 (11p15) and belongs to the cip/kip gene family. It encodes a cell cycle inhibitor that binds to G1 cyclin-CDK complexes. Thus p57KIP2 causes arrest of the cell cycle in G1 phase. CDKN1C was found to lead to cancer cell dormancy; its gene expression is regulated through the activity of glucocorticoid receptors (GRs) through chromatin remodelling mediated by SWI/SNF.

Background References

1. Lee JH et al. p57Kip2 imposes the reserve stem cell state of gastric chief cells. Cell Stem Cell. 2022 May

2. Creff J et al. Functional Versatility of the CDK Inhibitor p57Kip2. Front Cell Dev Biol. 2020 Oct

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P49918 Human

SwissProt: P49919 Mouse

Entrez Gene: 246060 Rat

Synonyms

Beckwith Wiedemann syndrome antibody

BWCR antibody

BWS antibody

CDKI antibody

CDKN 1C antibody

CDKN1C antibody

CDN1C_HUMAN antibody

Cyclin dependent kinase inhibitor 1C antibody

Cyclin dependent kinase inhibitor p57 antibody

Cyclin-dependent kinase inhibitor 1C antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of p57 Kip2 on different lysates with Rabbit anti-p57 Kip2 antibody (<a href="/products/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HeLa treated with 50nM dexamethasone for 16 hours cell lysate (20 µg/Lane)<br />Lane 3: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 4: K-562 cell lysate (20 µg/Lane)<br />Lane 5: 293T cell lysate (20 µg/Lane)<br />Lane 6: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 7: Rat brain tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 32 kDa<br />Observed band size: 50 kDa<br /><br />Exposure time: Lane 1-3: 59 seconds; Lane 4-7: 3 minutes; ECL: K1801;<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/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of p57 Kip2 on different lysates with Rabbit anti-p57 Kip2 antibody (HA722137) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HeLa treated with 50nM dexamethasone for 16 hours cell lysate (20 µg/Lane)
    Lane 3: SH-SY5Y cell lysate (20 µg/Lane)
    Lane 4: K-562 cell lysate (20 µg/Lane)
    Lane 5: 293T cell lysate (20 µg/Lane)
    Lane 6: Mouse brain tissue lysate (40 µg/Lane)
    Lane 7: Rat brain tissue lysate (40 µg/Lane)

    Predicted band size: 32 kDa
    Observed band size: 50 kDa

    Exposure time: Lane 1-3: 59 seconds; Lane 4-7: 3 minutes; ECL: K1801;
    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 (HA722137) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of p57 Kip2 on different lysates with Rabbit anti-p57 Kip2 antibody (<a href="/products/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 50nM dexamethasone for 16 hours cell lysate<br />Lane 3: HeLa treated with 50nM dexamethasone for 16 hours cell lysate, then the membrane treated with λpp for 1 hour<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 32 kDa<br />Observed band size: 50 kDa<br /><br />Exposure time: 59 seconds; ECL: K1801;<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/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of p57 Kip2 on different lysates with Rabbit anti-p57 Kip2 antibody (HA722137) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 50nM dexamethasone for 16 hours cell lysate
    Lane 3: HeLa treated with 50nM dexamethasone for 16 hours cell lysate, then the membrane treated with λpp for 1 hour

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 32 kDa
    Observed band size: 50 kDa

    Exposure time: 59 seconds; ECL: K1801;
    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 (HA722137) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HeLa cells treated with or without 50nM dexamethasone for 16 hours labeling p57 Kip2 with Rabbit anti-p57 Kip2 antibody (<a href="/products/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) at 1/250 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 Rabbit anti-p57 Kip2 antibody (<a href="/products/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) at 1/250 dilution in 1% BSA in PBST 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. 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/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution 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.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of HeLa cells treated with or without 50nM dexamethasone for 16 hours labeling p57 Kip2 with Rabbit anti-p57 Kip2 antibody (HA722137) at 1/250 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 Rabbit anti-p57 Kip2 antibody (HA722137) at 1/250 dilution in 1% BSA in PBST 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.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-p57 Kip2 antibody (<a href="/products/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) at 1/200 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/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) 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.

    Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-p57 Kip2 antibody (HA722137) at 1/200 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 (HA722137) 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.

  • Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-p57 Kip2 antibody (<a href="/products/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) at 1/200 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/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) 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.

    Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-p57 Kip2 antibody (HA722137) at 1/200 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 (HA722137) 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.

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-p57 Kip2 antibody (<a href="/products/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) at 1/200 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/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) 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.

    Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-p57 Kip2 antibody (HA722137) at 1/200 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 (HA722137) 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.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-p57 Kip2 antibody (<a href="/products/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) at 1/200 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/HA722137" style="font-weight: bold;text-decoration: underline;">HA722137</a>) 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.

    Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-p57 Kip2 antibody (HA722137) at 1/200 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 (HA722137) 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.

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