Human STAG2 (SA2) knockout HeLa cell line
- Advanced Validation
- What is this?
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STAG2 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: Insertion of the selection cassette in exon 11. 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
Cohesin subunit SA-2, DKFZp686P168, DKFZp781H1753, FLJ25871, SCC3 homolog 2, SCC3B, STAG2_HUMAN, Stromal antigen 2, bA517O1.1
- WB
Lab
Western blot - Human STAG2 (SA2) knockout HeLa cell line (AB265461)
Lanes 1-3 : Merged signal (red and green). Green - ab155081 observed at 141 kDa. Red - loading control ab8245 observed at 36 kDa.
ab155081 Anti-SA2 antibody [EPR10994(B)] was shown to specifically react with SA2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265461 (knockout cell lysate ab257707) was used. Wild-type and SA2 knockout samples were subjected to SDS-PAGE. ab155081 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 500 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-SA2 antibody [EPR10994(B)] (<a href='/en-us/products/primary-antibodies/sa2-antibody-epr10994b-ab155081'>ab155081</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
STAG2 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human STAG2 (SA2) knockout HeLa cell line (ab265461)
Lane 3:
HCT116 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: 141 kDa
Observed band size: 141 kDa
false
- WB
Lab
Western blot - Human STAG2 (SA2) knockout HeLa cell line (AB265461)
Lanes 1-3 : Merged signal (red and green). Green - ab201451 observed at 141 kDa. Red - loading control ab8245 observed at 36 kDa.
ab201451 Anti-SA2 antibody [EPR17865] - C-terminal was shown to specifically react with SA2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265461 (knockout cell lysate ab257707) was used. Wild-type and SA2 knockout samples were subjected to SDS-PAGE. ab201451 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (<a href='/en-us/products/primary-antibodies/sa2-antibody-epr17865-c-terminal-ab201451'>ab201451</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
STAG2 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human STAG2 (SA2) knockout HeLa cell line (ab265461)
Lane 3:
HCT116 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: 141 kDa
Observed band size: 141 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human STAG2 (SA2) knockout HeLa cell line (AB265461)
Homozygous : Insertion of the selection cassette in exon 11.
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
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
SA2 plays an important role in regulating gene expression by forming loops that connect different genomic regions. SA2 as a member of the cohesin complex also participates in DNA repair by facilitating accurate recombination enhancing genome stability. This involvement is essential for maintaining chromatin structure which influences diverse cellular processes. Its expression can be an important modulator in the dynamic environment of cellular proliferation and differentiation.
Pathways
SA2 functions integrally within the cohesin-mediated sister chromatid cohesion pathway and the DNA damage response pathway. In these pathways SA2 teams with proteins such as CTCF in insulator function regulation and BRCA1 in DNA repair mechanisms. The cohesin-SA2 interaction with these proteins ensures proper chromosomal architecture and integrity which are essential for the accurate transmission of genetic material across generations of cells.
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
AB201451
Anti-SA2 antibody [EPR17865] - C-terminal
RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (AB201451)](https://content.abcam.com/products/images/sa2-antibody-epr17865-c-terminal-ab201451--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img36577.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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