BRPF1 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: 34 bp deletion in exon 2.
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, Homozygous: 34 bp deletion in exon 2
Alternative names
BR140, BRPF1_HUMAN, Bromodomain and PHD finger containing 1, Bromodomain and PHD finger-containing protein 1, Peregrin, Protein Br140, bromodomain-containing protein, 140kD
Recommended products
BRPF1 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: 34 bp deletion in exon 2.
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, Homozygous: 34 bp deletion in exon 2
- Concentration
Properties
- Gene name
- BRPF1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
- Zygosity
- Homozygous
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Peregrin/BRPF1 also known as Peregrins is a protein with a molecular mass around 130 kDa. It functions primarily as a component of the MOZ/MORF histone acetyltransferase (HAT) complex. Peregrin/BRPF1 plays a mechanical role in the regulation of gene expression by altering chromatin structure through histone modification. It is expressed in various tissues with higher expression in the brain and testis indicating its importance in neurological and reproductive functions.
- Biological function summary
Peregrin/BRPF1 is a part of the MOZ/MORF complex which is important for acetylating histone H3 at lysine 23 influencing transcriptional activation. This action aids in transcription regulation and chromatin remodeling facilitating access of transcription factors to DNA. By participating in these processes Peregrin/BRPF1 helps maintain proper gene expression patterns essential for normal cell function and development.
- Pathways
BRPF1 has roles in processes like the cell cycle and embryonic development pathways because its histone acetylation influences gene expression. It interacts with other acetyltransferases such as EP300 and CREBBP which play key roles in transcription regulation and chromatin remodeling within these pathways. These interactions highlight BRPF1's critical involvement in coordinating cellular events and developmental processes.
- Associated diseases and disorders
Dysregulation of BRPF1 is associated with neurological disorders such as intellectual disability and specific cancers due to its role in chromatin remodeling and gene expression. In the context of intellectual disability it interacts with other proteins involved in chromatin structure like NSD1 and KDM6A. These connections illustrate the significant impact of Peregrin/BRPF1's regulation and its influence on health when its function becomes impaired.
Product promise
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4 product images
Western blot - Human BRPF1 (Peregrin) knockout HEK-293T cell line (ab266549)
False colour image of Western blot: Anti-Peregrin/BRPF1 antibody [EPR24069-57] staining at 1/500 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-Peregrin/BRPF1 antibody [EPR24069-57] ab259840 was shown to bind specifically to Peregrin/BRPF1. A band was observed at 120 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in BRPF1 knockout cell line ab266549 (knockout cell lysate Human BRPF1 (Peregrin) knockout HEK-293T cell lysate ab258795). To generate this image wild-type and BRPF1 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 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. 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.
Lanes 1, 2, 3, 5 and 6: Western blot - Anti-Peregrin/BRPF1 antibody [EPR24069-57] (Anti-Peregrin/BRPF1 antibody [EPR24069-57] ab259840) at 1/500 dilution
Lane 4: Western blot - Anti-Peregrin/BRPF1 antibody [EPR24069-57] (Anti-Peregrin/BRPF1 antibody [EPR24069-57] ab259840)
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lanes 2 - 3: BRPF1 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human BRPF1 (Peregrin) knockout HEK-293T cell line (ab266549)
Lane 4: Empty
Lane 5: PC-3 cell lysate at 20 µg
Lane 6: HepG2 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 137 kDa
Observed band size: 120 kDa
Western blot - Human BRPF1 (Peregrin) knockout HEK-293T cell line (ab266549)
False colour image of Western blot: Anti-Peregrin/BRPF1 antibody - N-terminal staining at 1/500 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-Peregrin/BRPF1 antibody - N-terminal ab226909 was shown to bind specifically to Peregrin/BRPF1. A band was observed at 160 145 and 138 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in BRPF1 knockout cell line ab266549 (knockout cell lysate Human BRPF1 (Peregrin) knockout HEK-293T cell lysate ab258795). To generate this image wild-type and BRPF1 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 °. 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.
Observed bands - 160 145 and 138 kDa
All lanes: Western blot - Anti-Peregrin/BRPF1 antibody - N-terminal (Anti-Peregrin/BRPF1 antibody - N-terminal ab226909) at 1/500 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: BRPF1 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human BRPF1 (Peregrin) knockout HEK-293T cell lysate (Human BRPF1 (Peregrin) knockout HEK-293T cell lysate ab258795)
Lane 2: Western blot - Human BRPF1 (Peregrin) knockout HEK-293T cell line (ab266549)
Lane 3: PC-3 cell lysate at 20 µg
Lane 4: HepG2 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 137 kDa
Cell Culture - Human BRPF1 (Peregrin) knockout HEK-293T cell line (ab266549)
Representative images BRPF1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
Sanger Sequencing - Human BRPF1 (Peregrin) knockout HEK-293T cell line (ab266549)
Homozygous: 34 bp deletion in exon 2
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