Human SCD (SCD1) knockout HeLa cell line (ab265220)

SCD KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 3 and Insertion of the selection cassette in exon 3.

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Images

Western blot - Human SCD (SCD1) knockout HeLa cell line (AB265220), expandable thumbnail

Publications

Key facts

Cell type: HeLa
Species or organism: Human
Tissue: Cervix
Form: Liquid
Knockout validation: Sanger Sequencing, Western blot
Mutation description: Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 3 and Insertion of the selection cassette in exon 3

Alternative names

ACOD_HUMAN, Acyl-CoA desaturase, Delta(9)-desaturase, Delta-9-Desaturase, FADS5, Fatty acid desaturase, PRO0998, Stearoyl-CoA desaturase

Key facts

Cell type: HeLa
Species or organism: Human
Tissue: Cervix
Form: Liquid
Knockout validation: Sanger Sequencing, Western blot
Mutation description: Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 3 and Insertion of the selection cassette in exon 3
Disease: Adenocarcinoma
Concentration

Properties

Gene name: SCD
Gene editing type: Knockout
Gene editing method: CRISPR technology
Knockout validation: Sanger Sequencing, Western blot

Quality control

STR analysis: CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level: EU: 2 US: 2
Adherent/suspension: Adherent
Gender: Female

Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO₂. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x10⁴ cells/cm². Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines

All seeding densities should be based on cell counts gained by established methods.
A guide seeding density of 2x10⁴ cells/cm² is recommended.
Cells should be passaged when they have achieved 80-90% confluence.

Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Storage

Shipped at conditions: Dry Ice
Appropriate short-term storage conditions: -196°C
Appropriate long-term storage conditions: -196°C

Notes

Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

We will provide viable cells that proliferate on revival.

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Stearoyl-CoA desaturase-1 (SCD1) also known as delta-9-desaturase is an enzyme integral to the synthesis of monounsaturated fats. This enzyme converts saturated fatty acyl-CoAs to monounsaturated fatty acyl-CoAs by introducing a double bond between the ninth and tenth carbon atoms a process called desaturation. SCD1 has a molecular weight of approximately 45 kDa and expresses in a variety of tissues including the liver adipose tissue and heart.

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.

Associated diseases and disorders

SCD1 is implicated in conditions like obesity and metabolic syndrome. Altered SCD1 activity links to insulin resistance as seen in obesity by modulating lipid profiles that influence insulin sensitivity. SCD1 associates with proteins like peroxisome proliferator-activated receptor gamma (PPARγ) which regulates adipogenesis emphasizing its role in metabolic disorders.

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9 product images

Western blot - Human SCD (SCD1) knockout HeLa cell line (ab265220)
anes 1-3: Merged signal (red and green). Green - Anti-SCD1 antibody [EPR21963] ab236868. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 50 kDa.
Anti-SCD1 antibody [EPR21963] 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 Human SCD (SCD1) knockout HeLa cell lysate ab257658) was used. Wild-type and SCD1 knockout samples were subjected to SDS-PAGE. Anti-SCD1 antibody [EPR21963] ab236868 and Anti-tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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] (Anti-SCD1 antibody [EPR21963] ab236868) 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 (ab265220)
Lane 3: HepG2 cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 42 kDa
Western blot - Human SCD (SCD1) knockout HeLa cell line (ab265220)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-SCD1 antibody [CD.E10] ab19862 observed at 36 kDa. Red - loading control Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
Anti-SCD1 antibody [CD.E10] 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 ab265220 (SCD knockout cell lysate Human SCD (SCD1) knockout HeLa cell lysate ab257658). Wild-type and SCD knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-SCD1 antibody [CD.E10] ab19862 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4° 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 (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed 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] (Anti-SCD1 antibody [CD.E10] ab19862) 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 (ab265220)
Lane 3: HepG2 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 36 kDa
Sanger Sequencing - Human SCD (SCD1) knockout HeLa cell line (ab265220)
Allele-5: Insertion of the selection cassette in exon 3.
Sanger Sequencing - Human SCD (SCD1) knockout HeLa cell line (ab265220)
Allele-4: Insertion of the selection cassette in exon 3.
Sanger Sequencing - Human SCD (SCD1) knockout HeLa cell line (ab265220)
Allele-1: 4 bp deletion in exon 3.
Sanger Sequencing - Human SCD (SCD1) knockout HeLa cell line (ab265220)
Allele-2: 4 bp deletion in exon 3.
Western blot - Human SCD (SCD1) knockout HeLa cell line (ab265220)
Anti-SCD1 antibody [EPR21963] ab236868 was shown to react with SCD1 in wild-type HeLa cells in Western blot with loss of signal observed in SCD1 knockout cell line ab265220. Wild-type HeLa and SCD1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-SCD1 antibody [EPR21963] ab236868 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-SCD1 antibody [EPR21963] (Anti-SCD1 antibody [EPR21963] ab236868) at 1/1000 dilution
Lane 1: Wild-type HeLa lysate at 35 µg
Lane 2: SCD1 knock-out HeLa lysate at 35 µg
Sanger Sequencing - Human SCD (SCD1) knockout HeLa cell line (ab265220)
Allele-3: 4 bp deletion in exon 3.
Western blot - Human SCD (SCD1) knockout HeLa cell line (ab265220)
Anti-SCD1 antibody [CD.E10] ab19862 was shown to react with SCD1 in wild-type HeLa cells in Western blot with loss of signal observed in SCD1 knockout cell line ab265220. Wild-type HeLa and SCD1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-SCD1 antibody [CD.E10] ab19862 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-SCD1 antibody [CD.E10] (Anti-SCD1 antibody [CD.E10] ab19862) at 1/1000 dilution
Lane 1: Wild-type HeLa lysate at 35 µg
Lane 2: SCD1 knock-out HeLa lysate at 35 µg