Human GLDC knockout A549 cell line
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GLDC KO cell line available to order. KO validated. Free of charge wild type control available. 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
GCE, GCSP_HUMAN, GLDC, Glycine cleavage system P protein, Glycine decarboxylase, Glycine dehydrogenase (decarboxylating) mitochondrial, Glycine dehydrogenase [decarboxylating], mitochondrial, Gycine dehydrogenase (decarboxylating), HYGN1, MGC138198, MGC138200, NKH, glycine cleavage system protein P, glycine decarboxylase P protein
- WB
Lab
Western blot - Human GLDC knockout A549 cell line (AB300972)
Western blot : Rabbit Polyclonal to Glycine decarboxylase ab204087 staining at 0.4 µg/mL, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 110 kDa in Wild-type A549 cell lysates with no signal observed at this size in GLDC knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-Glycine decarboxylase antibody (<a href='/en-us/products/primary-antibodies/glycine-decarboxylase-antibody-ab204087'>ab204087</a>) at 0.4 µg/mL
Lane 1:
Wild-type A549 at 20 µg
Lane 2:
Western blot - Human GLDC knockout A549 cell line (ab300972) at 20 µg
Lane 3:
HepG2 at 20 µg
Lane 4:
HAP1 at 20 µg
Lane 5:
Raji at 20 µg
Secondary
Lanes 1 - 5:
Goat anti-Rabbit 800CW at 1/20000 dilution
Lanes 1 - 5:
Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 110 kDa
Observed band size: 110 kDa,76 kDa
false
Product details
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type A549 cell line (ab288558). 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 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.
Properties and storage information
Gene name
Gene editing type
Gene editing method
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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
Glycine decarboxylase functions in cellular metabolism and photorespiration. It is part of the glycine cleavage system complex which is responsible for breaking down glycine into CO2 NH3 and a methylene group. The methylene group then enters the folate pool and contributes to various one-carbon metabolism pathways. This process helps regulate cellular levels of glycine and serine both important for nucleotide and protein synthesis as well as maintaining the balance of oxidative stress within cells.
Pathways
Glycine decarboxylase plays an important role in the folate and photorespiratory pathways. In the folate pathway interactions with serine hydroxymethyltransferase allow for the conversion of serine to glycine and the transfer of one-carbon units needed for nucleic acid synthesis. In the photorespiratory pathway its activity helps recycle 2-phosphoglycolate in plant mitochondria minimizing energy loss. The protein dihydrolipoamide dehydrogenase part of the same enzyme complex assists in maintaining the cycle of oxidation and reduction necessary for these functions.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
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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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