Human CDKN1A knockout HCT116 cell lysate
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- 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='/en-us/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='/en-us/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 seq
Unknown
Sanger Sequencing - Human CDKN1A knockout HCT116 cell lysate (AB256870)
Homozygous : 1 bp insertion in exon2
Complementary Products
Primary Antibodies
AB109520
RabMAb|Recombinant|KO Validated|Lab Essentials|20ul selling size
![Western blot - Anti-p21 antibody [EPR362] (AB109520)](https://content.abcam.com/products/images/p21-antibody-epr362-ab109520--western-blot-img422025.jpg)
- 4
- 16
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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