AB290614

Human PSEN1 knockout PC-3 cell line

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PSEN1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout.

View Alternative Names

AD3, Ad3h, FAD, Homo Sapiens Clone CC44 Senilin 1, PS-1, PS1-CTF12, PSEN1, PSN1_HUMAN, PSNL1, Presenilin-1 CTF12, Protein S182, S182

3 Images

Lab

Western blot - Human PSEN1 knockout PC-3 cell line (AB290614)

Western blot : Anti-PSEN1 antibody [5E12] (ab252856) staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab252856 was shown to bind specifically to PSEN1. A band was observed at 100 kDa in wild-type PC-3 cell lysates with no signal observed at this size in PSEN1 knockout cell line. To generate this image, wild-type and PSEN1 knockout PC-3 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-Rat IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Presenilin 1/PS-1 antibody [5E12] (<a href='/en-us/products/primary-antibodies/presenilin-1-ps-1-antibody-5e12-ab252856'>ab252856</a>) at 1/1000 dilution

Lane 1:

Wild-type PC-3 cell lysate at 20 µg

Lane 2:

PSEN1 knockout PC-3 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rat IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Observed band size: 100 kDa

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Lab

Western blot - Human PSEN1 knockout PC-3 cell line (AB290614)

Western blot : Anti-PSEN1 antibody [EP2000Y] (ab76083) staining at 1/2500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab76083 was shown to bind specifically to PSEN1. A band was observed at 18 kDa in wild-type PC-3 cell lysates with no signal observed at this size in PSEN1 knockout cell line. To generate this image, wild-type and PSEN1 knockout PC-3 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Presenilin 1/PS-1 antibody [EP2000Y] (<a href='/en-us/products/primary-antibodies/presenilin-1-ps-1-antibody-ep2000y-ab76083'>ab76083</a>) at 1/2500 dilution

Lane 1:

Wild-type PC-3 cell lysate at 20 µg

Lane 2:

PSEN1 knockout PC-3 cell lysate at 20 µg

Lane 3:

Wild-type HAP1 cell lysate at 20 µg

Lane 4:

PSEN1 knockout HAP1 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

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Supplier Data

Next Generation Sequencing - Human PSEN1 knockout PC-3 cell line (AB290614)

104 bp deletion after Tyr188 (allele 1) and 1 bp deletion after Tyr188 of the WT protein (allele 2).

Key facts

Cell type

PC-3

Species or organism

Human

Tissue

Prostate

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout

Disease

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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Product details

Recommended control: Human wild-type PC-3 cell line (ab290718). 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.

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What's included?

{ "values": { "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab290614-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab290614 Human PSEN1 knockout PC-3 cell line", "number":"AB290614-CMP01", "productcode":"" } ] } } }

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Properties and storage information

Gene name

PSEN1

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Next Generation Sequencing, Western blot

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

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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

F-12K + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Presenilin 1 often referred to as PS-1 is a protein with an important role in the function of gamma-secretase a multi-subunit protease complex. It weighs approximately 50 kDa and is widely expressed in different tissues especially in the brain. Presenilin 1 functions as part of the gamma-secretase complex which is responsible for processing several type I transmembrane proteins. This protein undergoes endoproteolytic processing to form an N-terminal and C-terminal fragment which are pivotal for its gamma-secretase activity.

Biological function summary

Presenilin 1 plays a fundamental role in the cleavage of amyloid precursor protein (APP) within the transmembrane domain. This cleavage results in the formation of amyloid-beta peptides which are of significant interest in neurobiology. Presenilin 1 is an integral component of the gamma-secretase complex that includes nicastrin APH-1 and PEN-2. This complex involvement underlines the enzyme's importance in cell signaling pathways and modulation of synaptic functions. The functionality at the synaptic level is particularly important given its expression pattern and involvement in neural processes.

Pathways

The action of presenilin 1 is closely linked to the Notch signaling pathway and the amyloidogenic pathway. Presenilin 1 facilitates the intramembrane cleavage of the Notch receptor releasing the Notch intracellular domain that translocates to the nucleus and affects gene transcription. The protein's role in processing APP into amyloid-beta peptides links it directly to Alzheimer's disease pathology. The protein Tax also a component of the gamma-secretase complex shows interdependencies in these pathways.

Presenilin 1 holds significant relevance in Alzheimer's disease where mutations in its gene are associated with early-onset familial Alzheimer's. The production of amyloid-beta peptides through presenilin 1's interaction with APP is central to the disease's pathogenesis. Another disorder linked to presenilin 1 is Dilated Cardiomyopathy where presenilin mutations may affect calcium signaling in cardiac cells. In Alzheimer's the interaction between presenilin 1 and the amyloid-beta peptide protein is critical while in cardiac issues it's more about protein misfolding and signaling mishaps.

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Target data

See full target information PSEN1

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

Human PSEN1 knockout PC-3 cell line

Western blot - Human PSEN1 knockout PC-3 cell line (AB290614)

Western blot - Human PSEN1 knockout PC-3 cell line (AB290614)

Next Generation Sequencing - Human PSEN1 knockout PC-3 cell line (AB290614)

Key facts

Cell type

Species or organism

Tissue

Form

Knockout validation

Mutation description

Disease

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Handling procedures

Initial handling guidelines

Subculture guidelines

Culture medium

Cryopreservation medium

Supplementary information

Biological function summary

Pathways

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Target data

Complementary Products

Cell Lines & Lysates

Cell Lines & Lysates

Primary Antibodies

Primary Antibodies

Product promise