Human GLA (Galactosidase alpha) knockout HeLa cell line
- Advanced Validation
- What is this?
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GLA 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 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
AGAL_HUMAN, Agalsidase alfa, Alpha D galactoside galactohydrolase 1, Alpha gal A, Alpha-D-galactosidase A, Alpha-D-galactoside galactohydrolase, Alpha-galactosidase A, GLA, GLA protein, Galactosidase, alpha, Melibiase
- WB
Lab
Western blot - Human GLA (Galactosidase alpha) knockout HeLa cell line (AB265563)
Lanes 1-2 : Merged signal (red and green). Green - ab168341 observed at 49 kDa. Red - loading control ab8245 observed at 37 kDa.
ab168341 Anti-Galactosidase alpha antibody [EP5828(2)] was shown to specifically react with Galactosidase alpha in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265563 (knockout cell lysate ab257449) was used. Wild-type and Galactosidase alpha knockout samples were subjected to SDS-PAGE. ab168341 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 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-Galactosidase alpha antibody [EP5828(2)] (<a href='/en-us/products/primary-antibodies/galactosidase-alpha-antibody-ep58282-ab168341'>ab168341</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
GLA knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human GLA (Galactosidase alpha) knockout HeLa cell line (ab265563)
Predicted band size: 38 kDa,49 kDa
Observed band size: 49 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human GLA (Galactosidase alpha) knockout HeLa cell line (AB265563)
Homozygous : 1 bp deletion 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 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?
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
The role of alpha-galactosidase involves the breakdown of complex carbohydrates particularly those containing alpha-galactosidic bonds. It does not operate as part of a complex but rather functions independently to cleave these specific bonds in glycolipids and glycoproteins. This activity is critical for degrading langerin and other gangliosides preventing the accumulation of these molecules in the lysosomes. By continuously performing this function alpha-galactosidase maintains glycosphingolipid homeostasis ensuring efficient cellular metabolism.
Pathways
The involvement of alpha-galactosidase in metabolic and lysosomal catabolic pathways is significant. One critical pathway is the glycolipid metabolism pathway where it acts alongside related enzymes such as beta-galactosidase. It also participates in the lysosomal degradation pathway working in conjunction with other lysosomal enzymes to prevent substrate buildup. The interaction with enzymes like lysosomal hydrolases ensures the proper breakdown of complex molecules into simpler ones that cells can utilize or excrete.
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
Complementary Products
Primary Antibodies
AB168341
Anti-Galactosidase alpha antibody [EP5828(2)]
RabMAb|Recombinant|KO Validated|20ul selling size|Trial Size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galactosidase alpha antibody [EP5828(2)] (AB168341)](https://content.abcam.com/products/images/galactosidase-alpha-antibody-ep58282-ab168341--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img91191.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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