PSENEN KO cell line available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 1 bp insertion; Frameshift: 100%.
Images
Key facts
- Cell type
HEK-293
- Species or organism
Human
- Tissue
Kidney
- Form
Liquid
- Knockout validation
Next Generation Sequencing, Western blot
Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 1 bp insertion; Frameshift: 100%
Alternative names
Gamma Secretase Subunit PEN2, Gamma-secretase subunit PEN-2, Hematopoietic stem/progenitor cells protein MDS033, MDS033, MSTP064, PEN2_HUMAN, PSEN2, Presenilin Enhancer 2, Presenilin enhancer 2 homolog, Presenilin enhancer protein 2, psenen
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PSENEN KO cell line available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 1 bp insertion; Frameshift: 100%.
Alternative names
Gamma Secretase Subunit PEN2, Gamma-secretase subunit PEN-2, Hematopoietic stem/progenitor cells protein MDS033, MDS033, MSTP064, PEN2_HUMAN, PSEN2, Presenilin Enhancer 2, Presenilin enhancer 2 homolog, Presenilin enhancer protein 2, psenen
Key facts
- Cell type
HEK-293
- Species or organism
Human
- Tissue
Kidney
- Form
Liquid
- Knockout validation
Next Generation Sequencing, Western blot
- Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 1 bp insertion; Frameshift: 100%
- Concentration
Properties
- Gene name
PSENEN
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Next Generation Sequencing, Western blot
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 long-term storage conditions
-196°C
Notes
Recommended control: Human wild-type HEK-293 cell line (Human wild-type HEK-293 cell line ab259776). 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, 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.
Supplementary info
- Activity summary
PEN2 also known as Presenilin Enhancer 2 is a component of the gamma-secretase complex. It has a molecular mass of approximately 12 kDa. PEN2 is expressed in a variety of tissues including neuronal cells and across several cell lines like the HEK 293. This protein plays an essential mechanical role in stabilizing and activating the gamma-secretase complex which is necessary for its protease activity.
- Biological function summary
PEN2 is an important element of the gamma-secretase complex composed of presenilin nicastrin APH-1 and PEN2. Its role involves the facilitation of proper maturation and assembly of the complex contributing to the proteolytic processing of several type I transmembrane proteins including the amyloid precursor protein (APP) and Notch. PEN2 plays a part in these cellular processes by ensuring that the gamma-secretase complex functions correctly impacting cellular signaling and communication.
- Pathways
Gamma-secretase activity where PEN2 is involved is central to the Notch signaling pathway and the amyloidogenic pathway. The Notch signaling pathway is fundamental in cell differentiation proliferation and apoptosis and relies significantly on the function of the gamma-secretase complex. The amyloidogenic pathway involves the processing of APP to produce amyloid-beta peptides which PEN2's role in the gamma-secretase complex influences. Related proteins in these pathways include presenilin which is an active component in Notch and APP cleavage.
- Associated diseases and disorders
Mutations or dysregulation of PEN2 can have links to Alzheimer's disease and certain cancers. Alzheimer's disease is associated with the production of amyloid-beta peptides which accumulate into plaques a process influenced by PEN2 through gamma-secretase activity with APP. Dysregulated Notch signaling where PEN2 is involved is linked to various cancers due to its role in controlling cell fate decisions. PEN2's interactions with proteins like presenilin and APP highlight its potential impact on these conditions.
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3 product images
Western blot - Human PSENEN (PEN2) knockout HEK-293 cell line (ab273720)
False colour image of Western blot: Anti-PEN2 antibody [EPR9200] staining at 1/1000 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-PEN2 antibody [EPR9200] ab154830 was shown to bind specifically to PEN2. A band was observed at 12 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in PSENEN knockout cell line ab273720 (knockout cell lysate Human PSENEN (PEN2) knockout HEK-293 cell lysate ab273772). To generate this image, wild-type and PSENEN knockout HEK-293 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 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-PEN2 antibody [EPR9200] (Anti-PEN2 antibody [EPR9200] ab154830) at 1/1000 dilution
Lane 1: Wild-type HEK-293 cell lysate at 20 µg
Lane 2: PSENEN knockout HEK-293 cell lysate at 20 µg
Lane 3: Neuro-2a cell lysate at 20 µg
Lane 4: Daudi cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 12 kDa
Observed band size: 12 kDa
Next Generation Sequencing - Human PSENEN (PEN2) knockout HEK-293 cell line (ab273720)
Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 1 bp insertion; Frameshift: 100%
Next Generation Sequencing - Human PSENEN (PEN2) knockout HEK-293 cell line (ab273720)
1 bp deletion (allele 1) and 1 bp insertion (allele 2) after Tyr18 of the WT protein
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