Human SPHK2 knockout HEK-293T cell lysate
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SPHK2 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 exon1.
View Alternative Names
SK 2, SPHK2_HUMAN, SPK 2, Sphingosine kinase 2, Sphingosine kinase type 2
- WB
Lab
Western blot - Human SPHK2 knockout HEK-293T cell lysate (AB258699)
Lane 1 : Wild-type HEK-293T cell lysate 20 μg
Lane 2 : SPHK2 knockout HEK-293T cell lysate 20 μg
Lane 3 : HeLa cell lysate 20 μg
Lane 4 : Raji cell lysate 20 μg
Lanes 1 - 4 : Merged signal (red and green). Green - Anti-SPHK2 antibody observed at 68 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-SPHK2 antibody was shown to react with SPHK2 in wild-type HEK-293T cells in Western blot with loss of signal observed in SPHK2 knockout cell line ab267293 (SPHK2 knockout cell lysate ab258699). Wild-type HEK-293T and SPHK2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with anti-SPHK2 antibody and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Anti-SPHK2 antibody at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
SPHK2 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human SPHK2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-sphk2-knockout-hek-293t-cell-line-ab267293'>ab267293</a>)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
Raji cell lysate at 20 µg
false
- Sanger seq
Unknown
Sanger Sequencing - Human SPHK2 knockout HEK-293T cell lysate (AB258699)
Homozygous : 1 bp insertion in exon1
- WB
Supplier Data
Western blot - Human SPHK2 knockout HEK-293T cell lysate (AB258699)
Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, ab320741 was shown to bind specifically to SPHK2. Target of interest was observed at 70 kDa in wild-type HEK293T cell lysate (Lane 1) with no signal observed at this size in SPHK2 knockout cell line (Lane 2) (Lane 2, knockout cell line ab267293/ knockout cell lysate ab258699). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes:
Western blot - Anti-SPHK2 antibody [EPR29189-33] (<a href='/en-us/products/primary-antibodies/sphk2-antibody-epr29189-33-ab320741'>ab320741</a>) at 1/1000 dilution
Lane 1:
Parental HEK293T(EDWT04) whole cell lysate at 20 µg
Lane 2:
Western blot - Human SPHK2 knockout HEK-293T cell lysate (ab258699) at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution
Observed band size: 70 kDa,36 kDa
false
Exposure time: 180s
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
SPHK2 influences cell growth survival and differentiation by generating sphingosine-1-phosphate a signaling lipid. It functions independently or in association with other proteins but it is not always a part of a larger complex. The generated S1P acts as a signaling molecule both inside and outside of cells affecting processes like apoptosis and inflammation.
Pathways
SPHK2 is a significant player in the sphingolipid signaling pathway and impacts the MAPK/ERK pathway. These pathways collectively regulate cellular proliferation and apoptosis. SPHK2 works in conjunction with SPHK1 a closely related sphingosine kinase in these signaling routes. These kinases together help to balance the levels of sphingosine and S1P which is essential for maintaining cellular homeostasis.
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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