Human PUS1 knockout HEK-293T cell lysate
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- WB
Lab
Western blot - Human PUS1 knockout HEK-293T cell lysate (AB258158)
Lane 1 : Wild-type HEK-293T cell lysate (20µg)
Lane 2 : PUS1 knockout HEK-293T cell lysate (20µg)
Lane 3 : HeLa cell lysate (20µg)
Lane 4 : Daudi cell lysate (20µg)
Lanes 1- 4 : Merged signal (red and green). Green - ab203010 observed at 45 kDa. Red - loading control ab7291 observed at 50 kDa.
ab203010 Anti-PUS1 antibody [EPR20181] was shown to specifically react with PUS1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266091 (knockout cell lysate ab258158) was used. Wild-type and PUS1 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. ab203010 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-PUS1 antibody [EPR20181] (<a href='/en-us/products/primary-antibodies/pus1-antibody-epr20181-ab203010'>ab203010</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
PUS1 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human PUS1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-pus1-knockout-hek-293t-cell-line-ab266091'>ab266091</a>)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Predicted band size: 47 kDa
Observed band size: 45 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PUS1 knockout HEK-293T cell lysate (AB258158)
Allele-2 : 41 bp deletion in exon2
- Sanger seq
Unknown
Sanger Sequencing - Human PUS1 knockout HEK-293T cell lysate (AB258158)
Allele-1 : 47 bp deletion in exon2
Complementary Products
Primary Antibodies
AB203010
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunocytochemistry/ Immunofluorescence - Anti-PUS1 antibody [EPR20181] (AB203010)](https://content.abcam.com/products/images/pus1-antibody-epr20181-ab203010--immunocytochemistry-immunofluorescence-img69246.jpg)
- 0
- 0
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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
The modification of RNA by PUS1 improves the stability and folding of the RNA structure aiding its functionality. The enzyme is not part of a larger complex but acts independently to achieve its role in RNA modification. Pseudouridine created by PUS1 permits enhanced base stacking and hydrogen bonding which influences the translation process and the overall efficiency of protein synthesis. This modification is important for the proper functioning of tRNA and rRNA ensuring fidelity during protein translation.
Pathways
PUS1 plays an essential role in the RNA processing and modification pathways. Its activity integrates into pathways such as ribosome biogenesis and RNA quality control. Particularly it interacts with other RNA-binding proteins and enzymes like TRUB1 another pseudouridine synthase which also contributes to the maturation of structured RNAs. These pathways are vital for maintaining cellular homeostasis and responding to various physiological demands.
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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