STAG2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 11.
Cohesin subunit SA-2, DKFZp686P168, DKFZp781H1753, FLJ25871, SCC3 homolog 2, SCC3B, STAG2_HUMAN, Stromal antigen 2, bA517O1.1
STAG2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 11.
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.
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.
The SA2 protein also known as stromal antigen 2 is a component of the cohesin complex playing a significant role in chromosomal dynamics. With an approximate molecular weight of 152 kDa SA2 exhibits high expression in the nucleus of proliferating cells particularly during interphase. This cohesin component contributes critically to maintaining sister chromatid cohesion and ensuring proper segregation during cell division serving as a functional partner in the cohesin complex with other components like SMC1 SMC3 and RAD21.
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.
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.
SA2 mutations or dysregulations have been linked to Cornelia de Lange Syndrome (CdLS) and certain cancers. In CdLS SA2 mutations disrupt normal cohesin function affecting developmental processes. In cancer contexts abnormal SA2 function or expression can lead to improper DNA repair contributing to genomic instability and tumorigenesis. It is often studied in concert with other cohesin proteins and regulatory elements like NIPBL which provides insights into cohesin-related pathologies and potential therapeutic targets.
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Anti-SA2 antibody [EPR10994(B)] 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 Human STAG2 (SA2) knockout HeLa cell lysate ab257707) was used. Wild-type and SA2 knockout samples were subjected to SDS-PAGE. Anti-SA2 antibody [EPR10994(B)] ab155081 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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)] (Anti-SA2 antibody [EPR10994(B)] ab155081) 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
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 141 kDa
Observed band size: 141 kDa
Anti-SA2 antibody [EPR17865] - C-terminal 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 Human STAG2 (SA2) knockout HeLa cell lysate ab257707) was used. Wild-type and SA2 knockout samples were subjected to SDS-PAGE. Anti-SA2 antibody [EPR17865] - C-terminal ab201451 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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 (Anti-SA2 antibody [EPR17865] - C-terminal ab201451) 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
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 141 kDa
Observed band size: 141 kDa
Homozygous: Insertion of the selection cassette in exon 11.
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