SMARCA4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 7 bp deletion in exon 4.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 7 bp deletion in exon 4
Alternative names
ATP-dependent helicase SMARCA4, BAF 190, BAF190A, BRG1 protein, BRG1-associated factor 190A, BRM/SWI2 related gene 1, Brahma protein like 1, Global transcription activator homologous sequence, Homeotic gene regulator, MRD16, Mitotic growth and transcription activator, Nuclear protein GRB1, Protein BRG-1, Protein brahma homolog 1, RTPS2, SMARC A4, SMCA4_HUMAN, SNF2, SNF2 like 4, SNF2-beta, SNF2B, SNF2L4, SNF2LB, SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 4, SWI2, Sucrose nonfermenting like 4, Transcription activator BRG1, global transcription activator snf2l4, hSNF2b
Recommended products
SMARCA4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 7 bp deletion in exon 4.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 7 bp deletion in exon 4
- Concentration
Properties
- Gene name
- SMARCA4
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
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5 product images
Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)
False colour image of Western blot: Anti-BRG1 antibody [EPNCIR111A] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-BRG1 antibody [EPNCIR111A] ab110641 was shown to bind specifically to BRG1. A band was observed at 185 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SMARCA4 knockout cell line ab255432 (knockout cell lysate Human SMARCA4 (BRG1) knockout HEK-293T cell lysate ab263853). To generate this image, wild-type and SMARCA4 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-BRG1 antibody [EPNCIR111A] (Anti-BRG1 antibody [EPNCIR111A] ab110641) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: SMARCA4 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)
Performed under reducing conditions.
Predicted band size: 185 kDa
Observed band size: 185 kDa
Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)
Lanes 1- 2: Merged signal (red and green). Green - Anti-BRG1 antibody [EPNCIR111A] ab110641 observed at 185 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) observed at 50 kDa.
Anti-BRG1 antibody [EPNCIR111A] ab110641 was shown to react with SMARCA4 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255432 (knockout cell lysate Human SMARCA4 (BRG1) knockout HEK-293T cell lysate ab263853) was used. Wild-type HEK-293T and SMARCA4 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-BRG1 antibody [EPNCIR111A] ab110641 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) overnight at 4°C at a 1 in 10000 dilution and a 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-BRG1 antibody [EPNCIR111A] (Anti-BRG1 antibody [EPNCIR111A] ab110641) at 1/10000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: SMARCA4 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)
Performed under reducing conditions.
Predicted band size: 185 kDa
Observed band size: 185 kDa
Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)
Lanes 1- 2: Merged signal (red and green). Green - Anti-BRG1 antibody [EPR3912] ab108318 observed at 185 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.
Anti-BRG1 antibody [EPR3912] ab108318 was shown to react with SMARCA4 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255432 (knockout cell lysate Human SMARCA4 (BRG1) knockout HEK-293T cell lysate ab263853) was used. Wild-type HEK-293T and SMARCA4 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-BRG1 antibody [EPR3912] ab108318 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 10000 dilution and a 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-BRG1 antibody [EPR3912] (Anti-BRG1 antibody [EPR3912] ab108318) at 1/10000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: SMARCA4 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)
Performed under reducing conditions.
Predicted band size: 185 kDa
Observed band size: 185 kDa
Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)
Allele-1: 7 bp deletion in exon 4
Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)
Allele-2: 1 bp deletion in exon 4.
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