Human GMNN (Geminin) knockout HeLa cell lysate
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- WB
Lab
Western blot - Human GMNN (Geminin) knockout HeLa cell lysate (AB257450)
Lane 1 : Wild-type HeLa cell lysate (20 μg)
Lane 2 : GMNN knockout HeLa cell lysate (20 μg)
Lane 3 : HepG2 cell lysate (20 μg)
Lane 4 : U-87 MG cell lysate (20 μg)
Lanes 1-4 : Merged signal (red and green). Green - ab195047 observed at 32 kDa. Red - loading control ab7291 observed at 50 kDa.
ab195047 Anti-Geminin antibody [EPR14637] was shown to specifically react with Geminin DNA Replication Inhibitor in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264972 (knockout cell lysate ab257450) was used. Wild-type and Geminin DNA Replication Inhibitor knockout samples were subjected to SDS-PAGE. ab195047 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 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-Geminin antibody [EPR14637] (<a href='/en-us/products/primary-antibodies/geminin-antibody-epr14637-ab195047'>ab195047</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa lysate at 20 µg
Lane 2:
Geminin DNA Replication Inhibitor knockout HeLa lysate at 20 µg
Lane 2:
Western blot - Human GMNN (Geminin) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-gmnn-geminin-knockout-hela-cell-line-ab264972'>ab264972</a>)
Lane 3:
HepG2 lysate at 20 µg
Lane 4:
U-87 MG lysate at 20 µg
Predicted band size: 24 kDa
Observed band size: 32 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human GMNN (Geminin) knockout HeLa cell lysate (AB257450)
Allele-1 : 11 bp deletion in exon 3
- Sanger seq
Lab
Sanger Sequencing - Human GMNN (Geminin) knockout HeLa cell lysate (AB257450)
Allele-2 : Insertion of the selection cassette in exon 3
Complementary Products
![Western blot - Anti-Geminin antibody [EPR14637] (AB195047)](https://content.abcam.com/products/images/geminin-antibody-epr14637-ab195047--western-blot-img2122.jpg)
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
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
In a cellular context Geminin forms part of a complex with the nuclear protein Cdt1. This complex is essential in the regulation of the cell cycle's S phase. Geminin's function restricts DNA replication by binding to Cdt1 and stopping its activity which is necessary during the G1/S phase transition. By doing so it preserves the integrity of genetic information and prevents re-replication. Geminin's inhibition remains strict until it is degraded during mitosis allowing Cdt1 to resume its function in the following cell cycle.
Pathways
The regulation of Geminin is important to maintaining proper DNA replication and cell cycle integrity. Geminin plays a role in the cell cycle control pathway particularly during the G1 to S phase transition. Additionally it interfaces with other proteins like Paxillin which is involved in cell adhesion and movement. Although not directly linked in a pathway Paxillin's role in regulating cellular dynamics can influence cell cycle processes in which Geminin is involved. Geminin also serves as a K blocker relevant in maintaining genetic stability.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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