APBB1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 4 bp deletion in exon 2.
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, Homozygous: 4 bp deletion in exon 2
Alternative names
APBB1_HUMAN, Adaptor protein FE65a2, Amyloid beta (A4) precursor protein binding family B member 1, Amyloid beta A4 precursor protein binding family B, Amyloid beta A4 precursor protein-binding family B member 1, Amyloid beta precursor protein binding family B member 1, Fe65 protein, Protein Fe65, RIR, stat like protein
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APBB1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 4 bp deletion in exon 2.
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, Homozygous: 4 bp deletion in exon 2
- Concentration
Properties
- Gene name
- APBB1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- 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
FE65 also known as APBB1 is a protein with a molecular mass of approximately 65 kDa. It belongs to the family of adaptor proteins with important roles in different cellular processes. This protein is expressed abundantly in the brain particularly in neurons where it interacts with other proteins to mediate signaling pathways. FE65 contains several conserved domains including a WW domain and two phosphotyrosine-binding (PTB) domains which facilitate its interaction with multiple partners.
- Biological function summary
FE65 plays a significant role in neuronal development and cognitive functions. It is part of a protein complex that includes the amyloid precursor protein (APP). This complex is important for the regulation of gene expression and cellular communication. FE65 also affects the nuclear signaling by interacting with the chromatin-remodeling complex. These interactions suggest that FE65 has a hand in influencing transcriptional activities within the cell.
- Pathways
FE65 is involved in the APP processing pathway and the development of Alzheimer's disease. It interacts with APP and influences its role in the cleavage and formation of amyloid-beta peptides. These peptides are key factors within the Alzheimer's disease pathway. Moreover FE65 connects with other proteins such as Abl which is involved in cell differentiation and migration processes. Through these pathways FE65 contributes to a broad range of neuronal functions and activities.
- Associated diseases and disorders
FE65 has a close connection to Alzheimer’s disease and cognitive impairments. In Alzheimer's disease FE65 interacts with APP modulating the production of amyloid-beta an important pathological component of the disease. The accumulation of amyloid-beta is linked to the progression of symptoms in Alzheimer’s patients. Additionally FE65 is associated with other proteins related to neuronal dysfunction and synaptic plasticity which may impact cognitive disorders.
Product promise
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3 product images
Western blot - Human APBB1 (FE65) knockout HEK-293T cell line (ab267294)
False colour image of Western blot: Anti-FE65 antibody [EPR3538] staining at 1/2000 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 red. In Western blot, Anti-FE65 antibody [EPR3538] ab91650 was shown to bind specifically to FE65. A band was observed at 65/85 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in APBB1 knockout cell line ab267294 (knockout cell lysate Human APBB1 (FE65) knockout HEK-293T cell lysate ab257833). To generate this image, wild-type and APBB1 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°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-FE65 antibody [EPR3538] (Anti-FE65 antibody [EPR3538] ab91650) at 1/2000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: APBB1 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human APBB1 (FE65) knockout HEK-293T cell line (ab267294)
Lane 3: SH-SY5Y cell lysate at 20 µg
Lane 4: A431 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 77 kDa
Observed band size: 65 kDa
Cell Culture - Human APBB1 (FE65) knockout HEK-293T cell line (ab267294)
Representative images of APBB1 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 XL Core microscope.
Sanger Sequencing - Human APBB1 (FE65) knockout HEK-293T cell line (ab267294)
Homozygous: 4 bp deletion in exon2
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