Human GBA knockout HeLa cell line
- Advanced Validation
- What is this?
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Human GBA knockout HeLa cell line available to order. Recommended control: Human wild-type HeLa cell line (ab255448).
View Alternative Names
Acid beta-glucosidase, Alglucerase, BETA GLUCOSIDASE, ACID, Beta-glucocerebrosidase, D-glucosyl-N-acylsphingosine glucohydrolase, EC 3.2.1.45, GBA1, GC-B, GCase, GLCM_HUMAN, GLUCOCEREBROSIDASE PSEUDOGENE, Gba protein, Glucocerebrosidase, Glucocerebrosidase (alt.), Glucosidase beta, Glucosidase, beta, acid, Glucosidase, beta; acid (includes glucosylceramidase), Glucosylceramidase, Imiglucerase, Lysosomal glucocerebrosidase, OTTHUMP00000033992, OTTHUMP00000033993, betaGC
- sELISA
Supplier Data
Sandwich ELISA - Human GBA knockout HeLa cell line (AB265038)
The concentration of GBA was measured in human wild type and knockout HeLa cell extracts based on a 12.5 µg/mL extract load. The mean concentration was determined to be 3,620 pg/mL in wild type extract and undetectable (ND) in knockout extract. The control and GBA knockout cell line extracts are available as ab265038.
- WB
Lab
Western blot - Human GBA knockout HeLa cell line (AB265038)
Lanes 1- 2 : Merged signal (red and green). Green - ab128879 observed at 60 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab128879 was shown to react with GBA in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265038 (knockout cell lysate ab256929) was used. Wild-type HeLa and GBA knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab128879 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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-GBA antibody [EPR5143(3)] (<a href='/en-us/products/primary-antibodies/gba-antibody-epr51433-ab128879'>ab128879</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
GBA knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human GBA knockout HeLa cell line (ab265038)
Predicted band size: 60 kDa
Observed band size: 60 kDa
false
- WB
Lab
Western blot - Human GBA knockout HeLa cell line (AB265038)
Lanes 1- 2 : Merged signal (red and green). Green - ab125065 observed at 70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab125065 was shown to react with GBA in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265038 (knockout cell lysate ab256929) was used. Wild-type HeLa and GBA knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab125065 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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-GBA antibody [EPR5142] (<a href='/en-us/products/primary-antibodies/gba-antibody-epr5142-ab125065'>ab125065</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
GBA knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human GBA knockout HeLa cell line (ab265038)
Predicted band size: 60 kDa
Observed band size: 70 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human GBA knockout HeLa cell line (AB265038)
Sequencing chromatogram displaying sequence edit in exon 3
- Cell Culture
Unknown
Cell Culture - Human GBA knockout HeLa cell line (AB265038)
Representative images of GBA knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human GBA knockout HeLa cell line (AB265038)
Homozygous : 1 bp insertion in exon 3.
Reactivity data
Product details
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
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
GBA plays an important role in sphingolipid metabolism. It participates in the degradation of glycolipids within the lysosome contributing to lipid recycling. It acts independently rather than as a part of a major enzymatic complex. Through its role in degrading glucosylceramide GBA influences cellular homeostasis and bioenergetics ensuring balance in neural and systemic lipid levels.
Pathways
GBA's enzymatic functions are integral to the glycosphingolipid metabolic pathway. It is involved in the downstream steps of the lysosomal degradation of glycosphingolipids. The pathway operates alongside other important proteins such as beta-glucosidase and CERT-related transfer proteins all of which contribute to membrane lipid organization and signal transduction processes.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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