BRD3 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.
Images
Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
Alternative names
BRD3_HUMAN, Bromodomain containing 3, Bromodomain-containing protein 3, KIAA0043, ORFX, Open reading frame X, RING3 like gene, RING3-like protein, RING3L
Recommended products
BRD3 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.
Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- BRD3
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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
- McCoY5a + 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
Recommended control: Human wild-type HCT116 cell line (ab288559). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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:
Terms & Conditions.
2 product images
Western blot - Human BRD3 knockout HCT116 cell line (ab287239)
Western blot: Anti-BRD3 antibody [EPR23743-226] (Anti-BRD3 antibody [EPR23743-226] ab300106) staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-BRD3 antibody [EPR23743-226] ab300106 was shown to bind specifically to BRD3. A band was observed at 90-100 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in BRD3 knockout cell line. To generate this image, wild-type and BRD3 knockout HCT 116 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 HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-BRD3 antibody [EPR23743-226] (Anti-BRD3 antibody [EPR23743-226] ab300106) at 1/500 dilution
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: Western blot - Human BRD3 knockout HCT116 cell line (ab287239)
Lane 2: BRD3 knockout HCT 116 cell lysate at 20 µg
Lane 3: Wild-type HEK-293T ab255553 cell lysate at 20 µg
Lane 4: BRD3 knockout HEK-293T ab260529 cell lysate at 20 µg
SecondaryLanes 1 - 4: HRP conjugated Goat anti-Rabbit (H+L) at 1/20000 dilution
Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 90-100 kDa
Next Generation Sequencing - Human BRD3 knockout HCT116 cell line (ab287239)
121 bp deletion (allele 1) and 43 bp deletion (allele 2) in exon 1, CCDS6980.1
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