Human CBS knockout HeLa cell line
- Advanced Validation
- What is this?
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CBSL KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 5 and 1 bp insertion in exon 5. 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
AI047524, AI303044, Beta-thionase, CBS_HUMAN, Cbs cystathionine beta-synthase, Cystathionine beta-synthase, EC 4.2.1.22, HIP 4, MGC18856, MGC18895, MGC37300, Methylcysteine synthase, OTTHUMP00000109416, OTTHUMP00000109418, Serine sulfhydrase
- sELISA
Supplier Data
Sandwich ELISA - Human CBS knockout HeLa cell line (AB264950)
Sandwich ELISA of ab312869 with the capture antibody dilution at 2 µg/mL and detector antibody dilution at 0.5 µg/mL. Interpolated concentrations of native CBS in human control wild type HeLa cell and CBS knockout HeLa cell based on 25 µg/mL extract loads. The concentrations of CBS were measured in duplicate and interpolated from the CBS standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CBS concentration was determined to be 4321.4 pg/mL in wild type HeLa extract (Human wild-type HeLa cell line ab255448) and undetectable in CBS knockout HeLa extract (Human CBS knockout HeLa cell line ab264950).
- WB
Lab
Western blot - Human CBS knockout HeLa cell line (AB264950)
Lanes 1-3 : Merged signal (red and green). Green - ab131155 observed at 70 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab131155 Anti-CBS antibody [EPR8578] was shown to specifically react with CBS in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264950 (knockout cell lysate ab257203) was used. Wild-type and CBS knockout samples were subjected to SDS-PAGE. ab131155 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-CBS antibody [EPR8578] (<a href='/en-us/products/primary-antibodies/cbs-antibody-epr8578-ab131155'>ab131155</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CBS knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CBS knockout HeLa cell line (ab264950)
Lane 3:
Raji cell lysate at 20 µg
Predicted band size: 61 kDa
Observed band size: 70 kDa
false
- WB
Lab
Western blot - Human CBS knockout HeLa cell line (AB264950)
Lanes 1-3 : Merged signal (red and green). Green - ab140600 observed at 70 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab140600 Anti-CBS antibody [EPR8579] was shown to specifically react with CBS in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264950 (knockout cell lysate ab257203) was used. Wild-type and CBS knockout samples were subjected to SDS-PAGE. ab140600 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-CBS antibody [EPR8579] (<a href='/en-us/products/primary-antibodies/cbs-antibody-epr8579-ab140600'>ab140600</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CBS knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CBS knockout HeLa cell line (ab264950)
Lane 3:
Raji cell lysate at 20 µg
Predicted band size: 61 kDa
Observed band size: 70 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CBS knockout HeLa cell line (AB264950)
Allele-2 : 1 bp insertion in exon 5.
- Cell Culture
Unknown
Cell Culture - Human CBS knockout HeLa cell line (AB264950)
Representative images of CBS 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 CBS knockout HeLa cell line (AB264950)
Allele-1 : 11 bp deletion in exon 5.
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
The enzyme CBS plays a role in the transsulfuration pathway where it helps in the metabolism of homocysteine. CBS is a part of a larger complex in some tissues where it interacts with other enzymes involved in sulfur amino acid metabolism. This function is important in maintaining cellular sulfur amino acid balance and protecting cells from oxidative stress caused by elevated levels of homocysteine.
Pathways
CBS functions in the transsulfuration pathway which connects the methionine cycle and the synthesis of glutathione. This pathway is essential for detoxification and antioxidant protection. CBS interrelates with other proteins like cystathionine gamma-lyase (CGL) within this pathway. The interplay between CBS and CGL ensures the conversion of homocysteine to cysteine which is a precursor for the synthesis of glutathione.
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
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBS antibody [EPR8579] (AB140600)](https://content.abcam.com/products/images/cbs-antibody-epr8579-ab140600--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img125476.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com