Human SCD (SCD1) knockout HeLa cell lysate
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(0 Publication)
- WB
Lab
Western blot - Human SCD (SCD1) knockout HeLa cell lysate (AB257658)
Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : SCD knockout HeLa cell lysate (20 ug)
Lane 3 : HepG2 cell lysate (20 ug)
Lanes 1-3 : Merged signal (red and green). Green - ab236868. Red - loading control ab8245 observed at 50 kDa.
ab236868 Anti-SCD1 antibody [EPR21963] was shown to specifically react with SCD1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265220 (knockout cell lysate ab257658) was used. Wild-type and SCD1 knockout samples were subjected to SDS-PAGE. ab236868 and Anti-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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-SCD1 antibody [EPR21963] (<a href='/en-us/products/primary-antibodies/scd1-antibody-epr21963-ab236868'>ab236868</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SCD knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SCD (SCD1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-scd-scd1-knockout-hela-cell-line-ab265220'>ab265220</a>)
Lane 3:
HepG2 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 1/10000 dilution
Predicted band size: 42 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SCD (SCD1) knockout HeLa cell lysate (AB257658)
Allele-1 : 4 bp deletion in exon3
- Sanger seq
Unknown
Sanger Sequencing - Human SCD (SCD1) knockout HeLa cell lysate (AB257658)
Allele-4 : Insertion of the selection cassette in exon3
- Sanger seq
Unknown
Sanger Sequencing - Human SCD (SCD1) knockout HeLa cell lysate (AB257658)
Allele-5 : Insertion of the selection cassette in exon3
- Sanger seq
Unknown
Sanger Sequencing - Human SCD (SCD1) knockout HeLa cell lysate (AB257658)
Allele-2 : 4 bp deletion in exon3
- Sanger seq
Unknown
Sanger Sequencing - Human SCD (SCD1) knockout HeLa cell lysate (AB257658)
Allele-3 : 4 bp deletion in exon3
- WB
Lab
Western blot - Human SCD (SCD1) knockout HeLa cell lysate (AB257658)
Lane 1 : Wild-type HeLa cell lysate 20 ug
Lane 2 : SCD knockout HeLa cell lysate 20 ug
Lane 3 : HepG2 cell lysate 20 ug
Lanes 1 - 3 : Merged signal (red and green). Green - ab19862 observed at 36 kDa. Red - loading control, ab2866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
ab19862 was shown to react with SCD1 in wild-type HeLa cells in western blot with loss of signal observed in SCD knockout cell line ab262177 (SCD knockout cell lysate ab257658). HeLa wild-type and SCD knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab19862 and ab2866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-SCD1 antibody [CD.E10] (<a href='/en-us/products/primary-antibodies/scd1-antibody-cde10-ab19862'>ab19862</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SCD (SCD1) knockout HeLa cell lysate (ab257658) at 20 µg
Lane 3:
HepG2 cell lysate at 20 µg
Predicted band size: 42 kDa
Observed band size: 36 kDa
false
Complementary Products
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
Quality control - STR analysis
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
SCD1 plays a significant role in fatty acid metabolism by regulating the balance of saturated and unsaturated fatty acids which impacts membrane fluidity and cellular signaling. SCD1 does not function as part of a multi-protein complex acting independently in the endoplasmic reticulum. Its expression and activity influence lipid biosynthesis and storage affecting energy homeostasis and lipid composition in cells.
Pathways
Fatty acid synthesis and oxidation pathways extensively involve SCD1. It participates mainly in the lipogenesis pathway interacting with proteins like acetyl-CoA carboxylase (ACC) and fatty acid synthase (FAS). SCD1 contributes to maintaining the ratio of oleate and palmitoleate key monounsaturated fatty acids important for cellular functions by integrating into the lipid metabolism network.
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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