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AB265388

Human GYS1 (Glycogen synthase 1) knockout HeLa cell line

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GYS1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

View Alternative Names

Glycogen synthase 1, GYS1, GYS

2 Images
Western blot - Human GYS1 (Glycogen synthase 1) knockout HeLa cell line (AB265388)
  • WB

Lab

Western blot - Human GYS1 (Glycogen synthase 1) knockout HeLa cell line (AB265388)

ab40810 Recombinant Anti-Glycogen synthase 1/GYS1 antibody [EP817Y] was shown to specifically react with Glycogen synthase 1/GYS1 in HeLa wild-type cells (ab255928). Loss of signal was observed when knockout cell line ab265388 (knockout cell lysate ab257462) was used. Wild-type and Glycogen synthase 1/GYS1 knockout samples were subjected to SDS-PAGE. ab40810 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 10000 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-Glycogen synthase 1/GYS1 antibody [EP817Y] (<a href='/en-us/products/primary-antibodies/glycogen-synthase-1-gys1-antibody-ep817y-ab40810'>ab40810</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa lysate at 20 µg

Lane 2:

Glycogen synthase 1/GYS1 knockout HeLa lysate at 20 µg

Lane 2:

Western blot - Human GYS1 (Glycogen synthase 1) knockout HeLa cell line (ab265388)

Lane 3:

HEK-293 lysate at 20 µg

Lane 4:

Daudi lysate at 20 µg

Predicted band size: 84 kDa

Observed band size: 80 kDa

false

Sanger Sequencing - Human GYS1 (Glycogen synthase 1) knockout HeLa cell line (AB265388)
  • Sanger seq

Unknown

Sanger Sequencing - Human GYS1 (Glycogen synthase 1) knockout HeLa cell line (AB265388)

Homozygous : 1 bp deletion in exon 1.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

We will provide viable cells that proliferate on revival.

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?

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Properties and storage information

Gene name
GYS1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Supplementary information

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

Glycogen synthase 1 (GYS1) also known as muscle glycogen synthase catalyzes the synthesis of glycogen from glucose. It adds glucose units from UDP-glucose to the non-reducing ends of glycogen chains facilitating glycogen elongation. The molecular mass of GYS1 is approximately 85 kDa. It is predominantly expressed in skeletal muscle tissue providing energy storage primarily in the form of glycogen. GYS1 is regulated through phosphorylation wherein it undergoes inactive to active conformational changes.
Biological function summary

This enzyme plays a critical role in the energy storage mechanism of muscle tissues. GYS1 functions as a part of the glycogen synthase complex interacting with various glycogen-associated proteins to modulate its activity. These interactions control the enzyme's affinity for its substrate and its susceptibility to regulatory signals like phosphorylation affecting glycogen synthesis efficiency. Through its activity GYS1 ensures a readily available reserve of glucose in muscle tissues for rapid energy production when required.

Pathways

Glycogen synthase 1 is a central component of the glycogen metabolism pathway. It acts alongside other enzymes such as glycogen phosphorylase which catalyzes glycogen breakdown. The balance between GYS1 and glycogen phosphorylase determines the rate of glycogen turnover within cells. GYS1 is further regulated by the insulin signaling pathway where insulin promotes glycogen synthesis by altering GYS1 activity alongside proteins like AKT influencing glucose homeostasis and energy management in tissues.

Dysregulation of GYS1 activity often associates with glycogen storage diseases notably Type 0 where deficiencies in glycogen storage lead to episodes of low blood sugar and reduced exercise capacity. Furthermore aberrations in GYS1 function contribute to insulin resistance and Type 2 diabetes conditions linked with impaired glucose metabolism. Altered GYS1 activity impacts proteins like glycogen phosphorylase affecting overall glycogen metabolism balance and contributing to metabolic disorders.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

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.

Product protocols

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