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AB256870

Human CDKN1A knockout HCT116 cell lysate

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CDKN1A KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon2.

대체 명칭 보기

CDK-interacting protein 1, SDI1, MDA-6, Melanoma differentiation-associated protein 6, CIP1, CDKN1, Cyclin-dependent kinase inhibitor 1, WAF1, MDA6, CDKN1A, CAP20, p21, PIC1

2 이미지
Western blot - Human CDKN1A knockout HCT116 cell lysate (AB256870)
  • WB

Lab

Western blot - Human CDKN1A knockout HCT116 cell lysate (AB256870)

Lane 1 : Wild-type HCT116 cell lysate (20μg)
Lane 2 : CDKN1A knockout HCT116 cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab109199 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.
ab109199 Anti-p21 antibody [EPR3993] was shown to specifically react with p21 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266860 (knockout cell lysate ab256870) was used. Wild-type and p21 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109199 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-p21 antibody [EPR3993] (<a href='/ko/products/primary-antibodies/p21-antibody-epr3993-ab109199'>ab109199</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 20 µg

Lane 2:

CDKN1A knockout HCT116 cell lysate at 20 µg

Lane 2:

Western blot - Human CDKN1A knockout HCT116 cell line (<a href='/ko/products/cell-lines/human-cdkn1a-knockout-hct116-cell-line-ab266860'>ab266860</a>)

Predicted band size: 18 kDa,197 kDa,36 kDa,47 kDa,72 kDa,84 kDa

Observed band size: 150 kDa,20 kDa,40 kDa,45 kDa,47 kDa

false

Sanger Sequencing - Human CDKN1A knockout HCT116 cell lysate (AB256870)
  • Sanger seq

Unknown

Sanger Sequencing - Human CDKN1A knockout HCT116 cell lysate (AB256870)

Homozygous : 1 bp insertion in exon2

주요 정보

세포 유형

HCT116

Species or organism

Human

조직

Colon

Knockout 검증

Sanger Sequencing,Western blot

Mutation 정보

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon2.

질병

Carcinoma

Reactivity 정보

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

제품 세부 정보

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

제품 구성

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab256870-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human wild-type HCT116 cell lysate", "number":"AB256870-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human CDKN1A knockout HCT116 cell lysate", "number":"AB256870-CMP01", "productcode":"" } ] } } }

특성 및 보관 정보

유전자명
CDKN1A
유전자 편집 종류
Knockout
유전자 편집 방식
CRISPR technology
Knockout 검증
Sanger Sequencing, Western blot
Zygosity
Homozygous
배송 시 보관 조건
Ambient - Can Ship with Ice
적절한 단기 보관 조건
-20°C
적절한 장기 보관 조건
-20°C

추가 정보

This supplementary information is collated from multiple sources and compiled automatically.

The p21 protein also known as CDKN1A is an important regulator of cell cycle progression. It acts as a cyclin-dependent kinase inhibitor where it binds to and inhibits the activity of cyclin-CDK2 or -CDK4 complexes. The molecular weight of p21 is approximately 21 kDa. Cellular expression of p21 is diverse; it occurs in various tissues reflecting its broad role in maintaining cell cycle control. This protein's expression level is often assessed using techniques such as p21 western blotting p21 ELISA and p21 IHC staining.
Biological function summary

P21 plays a critical role in cell cycle regulation and DNA damage response. It associates with the p53 tumor suppressor protein forming an essential part of the p53 signaling pathway under stress conditions. p21 arrests cells in G1 phase allowing DNA repair or leading to cellular senescence. Through these mechanisms p21 integrates signals from stress and damage emphasizing its importance as a mediator in cellular checkpoint control.

Pathways

P21 is closely involved in the p53 and the Transforming Growth Factor-beta (TGF-β) signaling pathways. This integration helps in transmitting growth-inhibitory signals. p21 interacts with proteins such as p53 and cyclins ensuring precise regulation of the cell cycle and growth arrest when necessary. Triggered by p53 activation p21 contributes to preventing undisturbed cell proliferation acting as a safeguard against tumor formation.

P21's deregulation associates with cancer and age-related diseases. Its impairment is a common feature in many cancers often linked with defects in the p53 pathway. Reduced p21 expression can contribute to unchecked cell division and tumor progression. Moreover in age-related diseases p21's role in promoting cellular senescence connects it with degenerative conditions. The intricate relationship between p21 and proteins such as p53 and cyclins highlights its potential as a therapeutic target for these disorders.

품질 관리

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

제품 프로토콜

Product promise

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