Human MLKL knockout HeLa cell lysate
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MLKL KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and 2 bp deletion in exon2.
View Alternative Names
9130019I15Rik, FLJ34389, MLKL_HUMAN, Mixed lineage kinase domain like, Mixed lineage kinase domain like pseudokinase, Mixed lineage kinase domain-like protein, hMLKL
- WB
Unknown
Western blot - Human MLKL knockout HeLa cell lysate (AB263788)
Lane 1 : HUVEC cell lysate (20 μg)
Lane 2 : HT-29 cell lysate (20 μg)
Lane 3 : Wild-type HeLa cell lysate (20 μg)
Lane 4 : MLKL knockout HeLa cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab184718 observed at 54 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab184718 was shown to react with MLKL in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255408 (knockout cell lysate ab263788) was used. Wild-type and MLKL knockout samples were subjected to SDS-PAGE. ab184718 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-MLKL antibody [EPR17514] (<a href='/en-us/products/primary-antibodies/mlkl-antibody-epr17514-ab184718'>ab184718</a>) at 1/1000 dilution
Lane 1:
HUVEC cell lysate at 20 µg
Lane 2:
HT-29 cell lysate at 20 µg
Lane 2:
Western blot - Human MLKL knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-mlkl-knockout-hela-cell-line-ab255408'>ab255408</a>)
Lane 3:
Wild-type HeLa cell lysate at 20 µg
Lane 4:
MLKL knockout HeLa cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 54 kDa
Observed band size: 37 kDa,54 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human MLKL knockout HeLa cell lysate (AB263788)
Allele-1 : 2 bp deletion in exon2
- Sanger seq
Unknown
Sanger Sequencing - Human MLKL knockout HeLa cell lysate (AB263788)
Allele-2 : 1 bp insertion in exon2
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
The MLKL protein acts as an executioner of cell death by forming a complex that disrupts the plasma membrane integrity. This process is downstream of receptor-interacting serine/threonine-protein kinase 3 (RIPK3) which phosphorylates MLKL to form the active necrosome complex. Active MLKL oligomerizes and migrates towards the inner leaflet of the plasma membrane binding to phosphatidylinositol phosphates which assists in pore formation and cellular rupture. The ability to measure MLKL activity levels such as via MLKL ELISA kits is important for understanding necrotic processes in detailed studies.
Pathways
MLKL is integrally involved in the necroptotic pathway alongside RIPK1 and RIPK3 which are key initiators of necroptosis. Phosphorylated MLKL acts downstream of RIPK3 resulting in cell death without caspase activation distinguishing necroptosis from apoptosis. MLKL and RIPK3 are tightly linked within this pathway with MLKL phosphorylation serving as a vital event for the execution phase. The necroptosis pathway is part of larger networks including inflammatory response pathways highlighting the importance of MLKL's role beyond sheer cell death.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
![Western blot - Anti-MLKL antibody [EPR17514] (AB184718)](https://content.abcam.com/products/images/mlkl-antibody-epr17514-ab184718--western-blot-img85391.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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