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p21 Recombinant Rabbit Monoclonal Antibody [PSH22-37]

Usd: 385

Specification

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Catalog# HA724295

p21 Recombinant Rabbit Monoclonal Antibody [PSH22-37]

Application

  • WB

  • IF-Cell

  • IHC-P

  • IP

Reactivity

  • Human

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

p21 Recombinant Rabbit Monoclonal Antibody [PSH22-37]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IHC-P, IP

Molecular Weight

Predicted band size: 18 kDa

Positive Control

HCT 116 cell lysate, A549 cell lysate, MCF7 cell lysate, MCF7 treated with 10μM Nutlin 3a for 24 hours cell lysate, MCF7, human colon cancer tissue, human ovarian cancer tissue, human breast cancer tissue.

Conjugation

unconjugated

Clone Number

PSH22-37

Product Features

Form

Liquid

Concentration

Storage Instructions

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

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:5,000

  • IHC-P

  • 1:200

  • IP

  • 1-2μg/sample

Target

Function

p21Cip1 (alternatively p21Waf1), also known as cyclin-dependent kinase inhibitor 1 or CDK-interacting protein 1, is a cyclin-dependent kinase inhibitor (CKI) that is capable of inhibiting all cyclin/CDK complexes, though is primarily associated with inhibition of CDK2. p21 represents a major target of p53 activity and thus is associated with linking DNA damage to cell cycle arrest. This protein is encoded by the CDKN1A gene located on chromosome 6 (6p21.2) in humans.

Background References

1. Engeland K. Cell cycle regulation: p53-p21-RB signaling. Cell Death Differ. 2022 May

2. Bautista L et al . p21-Activated Kinases in Thyroid Cancer. Endocrinology. 2020 Aug

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P38936 Human

Synonyms

CAP20 antibody

CDK-interacting protein 1 antibody

CDKI antibody

CDKN1 antibody

Cdkn1a antibody

CDN1A_HUMAN antibody

CIP1 antibody

Cyclin Dependent Kinase Inhibitor 1A antibody

Cyclin-dependent kinase inhibitor 1 antibody

Cyclin-dependent kinase inhibitor 1A (P21) antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE),Cell treatment (CT)</span><br /><br />Western blot analysis of p21 on different lysates with Rabbit anti-p21 antibody (<a href="/products/HA724295" style="font-weight: bold;text-decoration: underline;">HA724295</a>) at 1/2,000 dilution.<br /><br />Lane 1: HCT 116 (Human colon cancer cells)<br />Lane 2: NCI-H1299 (Human non-small cell Lung carcinoma cells) (Negative)<br />Lane 3: A549 (Human lung adenocarcinoma cells)<br />Lane 4: MCF7 (Human breast cancer cells)<br />Lane 5: MCF7 (Human breast cancer cells) treated with Nutlin 3a (10uM, 24h)<br /><br />Lysates/proteins at 10 µg/Lane.<br />Exposure time: 1 minutes; ECL: K1801<br /><br />H1299 is a negative control (PMID: 23268708).<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA724295" style="font-weight: bold;text-decoration: underline;">HA724295</a>, 1/2,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 21 kDa<br />Observed band size: 21 kDa

    ☑ Relative expression (RE),Cell treatment (CT)

    Western blot analysis of p21 on different lysates with Rabbit anti-p21 antibody (HA724295) at 1/2,000 dilution.

    Lane 1: HCT 116 (Human colon cancer cells)
    Lane 2: NCI-H1299 (Human non-small cell Lung carcinoma cells) (Negative)
    Lane 3: A549 (Human lung adenocarcinoma cells)
    Lane 4: MCF7 (Human breast cancer cells)
    Lane 5: MCF7 (Human breast cancer cells) treated with Nutlin 3a (10uM, 24h)

    Lysates/proteins at 10 µg/Lane.
    Exposure time: 1 minutes; ECL: K1801

    H1299 is a negative control (PMID: 23268708).

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724295, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 21 kDa
    Observed band size: 21 kDa

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: MCF7 (Human cervix adenocarcinoma epithelial cell) treated or untreated with Nutlin 3a (10 μM, 24h)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.<br /><br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br />Primary antibody: <a href="/products/HA724295" style="font-weight: bold;text-decoration: underline;">HA724295</a>, 1/5,000, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature.<br /><br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).

    ☑ Cell treatment (CT)

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: MCF7 (Human cervix adenocarcinoma epithelial cell) treated or untreated with Nutlin 3a (10 μM, 24h)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.

    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.
    Primary antibody: HA724295, 1/5,000, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

    Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Colon cancer<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× PBST<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/HA724295" style="font-weight: bold;text-decoration: underline;">HA724295</a>, 1/200, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Colon cancer
    Sample: Paraffin-embedded section

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

    Wash buffer: 1× PBST
    Endogenous peroxidase blocking: 3% H2O2, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724295, 1/200, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: 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× PBST<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/HA724295" style="font-weight: bold;text-decoration: underline;">HA724295</a>, 1/200, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Ovarian Carcinoma
    Sample: Paraffin-embedded section

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

    Wash buffer: 1× PBST
    Endogenous peroxidase blocking: 3% H2O2, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724295, 1/200, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Invasive Ductal Carcinoma of the Breast<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× PBST<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/HA724295" style="font-weight: bold;text-decoration: underline;">HA724295</a>, 1/200, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Invasive Ductal Carcinoma of the Breast
    Sample: Paraffin-embedded section

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

    Wash buffer: 1× PBST
    Endogenous peroxidase blocking: 3% H2O2, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724295, 1/200, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunoprecipitation (IP)<br /><br />p21 was immunoprecipitated in 0.2 mg HCT 116 (Human colon cancer cell) cell lysate with <a href="/products/HA724295" style="font-weight: bold;text-decoration: underline;">HA724295</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA724295" style="font-weight: bold;text-decoration: underline;">HA724295</a> at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HCT 116 cell lysate (input)<br />Lane 2: Rabbit IgG instead of <a href="/products/HA724295" style="font-weight: bold;text-decoration: underline;">HA724295</a> in HCT 116 cell lysate<br />Lane 3: <a href="/products/HA724295" style="font-weight: bold;text-decoration: underline;">HA724295</a> IP in HCT 116 cell lysate<br /><br />Exposure time: 1 minute<br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br /><br />Primary dilution: <a href="/products/HA724295" style="font-weight: bold;text-decoration: underline;">HA724295</a>, 1/2,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), 2 hours at room temperature<br /><br />Predicted band size: 21 kDa<br />Observed band size: 21 kDa

    Application: Immunoprecipitation (IP)

    p21 was immunoprecipitated in 0.2 mg HCT 116 (Human colon cancer cell) cell lysate with HA724295 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724295 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HCT 116 cell lysate (input)
    Lane 2: Rabbit IgG instead of HA724295 in HCT 116 cell lysate
    Lane 3: HA724295 IP in HCT 116 cell lysate

    Exposure time: 1 minute
    Blocking: 5% NFDM/TBST, 1 hour at room temperature

    Primary dilution: HA724295, 1/2,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

    Predicted band size: 21 kDa
    Observed band size: 21 kDa

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

Citation

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