SMARCA4 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. 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
- 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
What's included?
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SMARCA4 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. 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
- 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
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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.
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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 lysate (ab263853)
Lane 1: Wild-type HEK-293T cell lysate 20 μg
Lane 2: SMARCA4 knockout HEK-293T cell lysate 20 μg
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 Human SMARCA4 (BRG1) knockout HEK-293T cell line ab255432 (knockout 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 (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 lysate (ab263853)
Lane 1: Wild-type HEK-293T cell lysate (20 ug)
Lane 2: SMARCA4 knockout HEK-293T cell lysate (20 ug)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 Human SMARCA4 (BRG1) knockout HEK-293T cell line ab255432 (knockout 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 (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 lysate (ab263853)
Lane 1: Wild-type HEK-293T cell lysate (20µg)
Lane 2: SMARCA4 knockout HEK-293T cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - Anti-BRG1 antibody [EPR3912] ab108318 observed at 185 kDa. Red - loading control Anti-Vinculin antibody [VIN-54] ab130007 observed at 124 kDa.
Anti-BRG1 antibody [EPR3912] ab108318 Recombinant Anti-BRG1 antibody [EPR3912] was shown to specifically react with SMARCA4 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human SMARCA4 (BRG1) knockout HEK-293T cell line ab255432 (knockout cell lysate ab263853) was used. Wild-type and SMARCA4 knockout samples 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] were incubated overnight at 4°C at 1 in 10000 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-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 (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 lysate (ab263853)
Allele-1: 7 bp deletion in exon 4
Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK-293T cell lysate (ab263853)
Allele-2: 1 bp deletion in exon 4
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