Human GNS knockout HeLa cell line
- Advanced Validation
- What is this?
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GNS KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 1 bp insertion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
2610016K11Rik, AU042285, C87209, G6S, GNS_HUMAN, Glucosamine (N-acetyl) 6 sulfatase, Glucosamine-6-sulfatase, MGC21274, N acetylglucosamine 6 sulfatase [Precursor], N-acetylglucosamine-6-sulfatase, N28088
- WB
Lab
Western blot - Human GNS knockout HeLa cell line (AB265495)
Lanes 1-3 : Merged signal (red and green). Green - ab154177 observed at 90 kDa. Red - loading control ab7291 observed at 50 kDa.
ab154177 Anti-GNS antibody [EPR8329(2)] was shown to specifically react with GNS in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265495 (knockout cell lysate ab257975) was used. Wild-type and GNS knockout samples were subjected to SDS-PAGE. ab154177 and 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 (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-GNS antibody [EPR8329(2)] (<a href='/en-us/products/primary-antibodies/gns-antibody-epr83292-ab154177'>ab154177</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
GNS knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human GNS knockout HeLa cell line (ab265495)
Lane 3:
293T 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: 62 kDa
Observed band size: 90 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human GNS knockout HeLa cell line (AB265495)
Allele-1 : 1 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human GNS knockout HeLa cell line (AB265495)
Allele-2 : 1 bp insertion in exon 1.
Reactivity data
Product details
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.
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
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Glucosamine (N-acetyl)-6-sulfatase activity influences cellular homeostasis by acting within the lysosomal hydrolase family. GNS participates in the breakdown and metabolism of glycosaminoglycans important for maintaining cellular integrity and signaling functions. It operates as a standalone enzyme rather than part of a larger complex. Through its enzymatic function GNS helps prevent the accumulation of partially degraded molecules inside lysosomes.
Pathways
Glucosamine (N-acetyl)-6-sulfatase activity plays a role in lysosomal storage pathways and in the catabolic processes of heparan sulfate. This enzyme works alongside other lysosomal proteins like iduronate sulfatase and alpha-L-iduronidase which are also involved in glycosaminoglycan degradation pathways. Proper function of these pathways is important for cellular waste management and nutrient recycling.
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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