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AB266016

Human APH1A knockout HeLa cell line

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APH1A KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 1 bp insertion in exon 1 and 7 bp deletion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
3 Images
Sanger Sequencing - Human APH1A knockout HeLa cell line (AB266016)
  • Sanger seq

Unknown

Sanger Sequencing - Human APH1A knockout HeLa cell line (AB266016)

Allele-3 : 1 bp insertion in exon 1.

Sanger Sequencing - Human APH1A knockout HeLa cell line (AB266016)
  • Sanger seq

Unknown

Sanger Sequencing - Human APH1A knockout HeLa cell line (AB266016)

Allele-2 : 1 bp deletion in exon 1.

Sanger Sequencing - Human APH1A knockout HeLa cell line (AB266016)
  • Sanger seq

Unknown

Sanger Sequencing - Human APH1A knockout HeLa cell line (AB266016)

Allele-1 : 7 bp deletion in exon 1.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 1 bp insertion in exon 1 and 7 bp deletion in exon 1

Disease

Adenocarcinoma

Product details

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.

What's included?

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Properties and storage information

Gene name
APH1A
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

The APH1a protein also known as Anterior Pharynx-1a plays a mechanical role as a part of the gamma-secretase complex. It has a mass of approximately 29 kDa. APH1a is expressed in various tissues with a high presence in brain regions and other human tissues. As a gamma-secretase component APH1a contributes to the intramembrane proteolysis of type I transmembrane proteins such as the Notch receptor and amyloid precursor protein (APP).
Biological function summary

As part of the gamma-secretase complex the APH1a protein facilitates intra-membrane cleavage processes. It associates with other key complex members like presenilin nicastrin and PEN-2. This complex's activity is important for the proper functioning of the Notch signaling pathway and amyloid-beta production both of which have important roles in cell differentiation and signaling in various organisms.

Pathways

APH1a is directly implicated in both the Notch signaling and amyloid precursor protein pathways. These are significant pathways impacting cellular communication and plaque formation in the brain. APH1a's interaction with proteins such as presenilin and nicastrin highlights its broader regulatory role particularly influencing developmental processes through Notch signaling and contributing to amyloid-beta genesis in neurobiological contexts.

Researchers associate APH1a with neurodegenerative diseases such as Alzheimer's disease and cancer. In Alzheimer's its role in the gamma-secretase complex impacts amyloid-beta production a substance linked to the neurodegenerative process. Additionally alterations in the Notch signaling pathway involving APH1a and related proteins like presenilin may contribute to certain cancers due to its influence on cell proliferation and differentiation. Understanding APH1a's interactions and functions offers valuable insights into these disease mechanisms.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

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'.

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