Human APBB1 (FE65) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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APBB1 KO cell line available to order. KO validated by. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 4 bp deletion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View 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
- WB
Lab
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] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-FE65 antibody [EPR3538] (<a href='/en-us/products/primary-antibodies/fe65-antibody-epr3538-ab91650'>ab91650</a>) 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
Predicted band size: 77 kDa
Observed band size: 65 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human APBB1 (FE65) knockout HEK-293T cell line (AB267294)
Homozygous : 4 bp deletion in exon2
- Cell Culture
Unknown
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.
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com