BRD2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 7 and 1 bp insertion in exon 7 and 22 bp deletion in exon 7.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 7 and 1 bp insertion in exon 7 and 22 bp deletion in exon 7
Alternative names
BRD2_HUMAN, Bromodomain containing 2, Bromodomain-containing protein 2, D6S113E, DKFZp686N0336, FLJ31942, FSH, FSRG1, Female sterile homeotic related gene 1, Female sterile homeotic related gene 1, mouse, homolog of, KIAA9001, NAT, O27.1.1, RING3, RING3 PROTEIN, RNF3, Really interesting new gene 3 protein
Recommended products
BRD2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 7 and 1 bp insertion in exon 7 and 22 bp deletion in exon 7.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 7 and 1 bp insertion in exon 7 and 22 bp deletion in exon 7
- Concentration
Properties
- Gene name
- BRD2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
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, 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.
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5 product images
Western blot - Human BRD2 knockout HEK-293T cell line (ab267265)
Lanes 1-3: Merged signal (red and green). Green - Anti-BRD2 antibody [EPR7642] - ChIP Grade ab139690 observed at 110 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.Anti-BRD2 antibody [EPR7642] - ChIP Grade ab139690 Anti-BRD2 antibody [EPR7642] was shown to specifically react with BRD2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267265 (knockout cell lysate Human BRD2 knockout HEK-293T cell lysate ab257191) was used. Wild-type and BRD2 knockout samples were subjected to SDS-PAGE. Anti-BRD2 antibody [EPR7642] - ChIP Grade ab139690 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2.5 hours 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-BRD2 antibody [EPR7642] - ChIP Grade (Anti-BRD2 antibody [EPR7642] - ChIP Grade ab139690) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: BRD2 knockout HEK293T cell lysate at 20 µg
Lane 2: Western blot - Human BRD2 knockout HEK-293T cell line (ab267265)
Lane 3: HeLa cell lysate at 20 µg
SecondaryAll 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: 126 kDa, 19 kDa, 20 kDa, 26 kDa, 32 kDa, 34 kDa, 44 kDa, 88 kDa
Observed band size: 110 kDa, 26 kDa, 50 kDa
Western blot - Human BRD2 knockout HEK-293T cell line (ab267265)
Lanes 1-3: Merged signal (red and green). Green - Anti-BRD2 antibody [BL-167-2A2] ab243865 observed at 110 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.Anti-BRD2 antibody [BL-167-2A2] ab243865 Anti-BRD2 antibody [BL-167-2A2] was shown to specifically react with BRD2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267265 (knockout cell lysate Human BRD2 knockout HEK-293T cell lysate ab257191) was used. Wild-type and BRD2 knockout samples were subjected to SDS-PAGE. Anti-BRD2 antibody [BL-167-2A2] ab243865 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2. 5 hours 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-BRD2 antibody [BL-167-2A2] (Anti-BRD2 antibody [BL-167-2A2] ab243865) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: BRD2 knockout HEK293T cell lysate at 20 µg
Lane 2: Western blot - Human BRD2 knockout HEK-293T cell line (ab267265)
Lane 3: HeLa cell lysate at 20 µg
SecondaryAll 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: 88 kDa
Observed band size: 110 kDa
Sanger Sequencing - Human BRD2 knockout HEK-293T cell line (ab267265)
Allele-1: 22 bp deletion in exon7
Sanger Sequencing - Human BRD2 knockout HEK-293T cell line (ab267265)
Allele-3: 1 bp insertion in exon 7.
Sanger Sequencing - Human BRD2 knockout HEK-293T cell line (ab267265)
Allele-2: 1 bp deletion in exon 7.
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