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AB266182

Human AKAP1 knockout HEK-293T cell line

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AKAP1 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 2 and Insertion of the selection cassette in exon 2. 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 AKAP1 knockout HEK-293T cell line (AB266182)
  • Sanger seq

Unknown

Sanger Sequencing - Human AKAP1 knockout HEK-293T cell line (AB266182)

Allele-2 : Insertion of the selection cassette in exon 2.

Cell Culture - Human AKAP1 knockout HEK-293T cell line (AB266182)
  • Cell Culture

Unknown

Cell Culture - Human AKAP1 knockout HEK-293T cell line (AB266182)

Representative images of AKAP1 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 AKAP1 knockout HEK-293T cell line (AB266182)
  • Sanger seq

Unknown

Sanger Sequencing - Human AKAP1 knockout HEK-293T cell line (AB266182)

Allele-1 : 11 bp deletion in exon 2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 2 and Insertion of the selection cassette in exon 2

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
AKAP1
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.

A-kinase anchoring protein 1 (AKAP1) also known as AKAP121 or D-AKAP1 is a protein that serves as an anchor point for signaling molecules. AKAP1 is approximately 121 kDa in mass. It is an important player in coordinating cellular processes by anchoring protein kinase A (PKA) to specific cellular compartments such as mitochondria. The protein is mainly expressed in the heart liver and brain but also found in other tissues indicating its broad role in cellular function.
Biological function summary

AKAP1 connects with several components to influence cellular signaling pathways and energy metabolism. It is a part of a complex network that involves mitochondrial targeting sequences which guide proteins to correct mitochondrial compartments. AKAP1 helps regulate processes like mitochondrial dynamics apoptosis and oxidative phosphorylation. These functions ensure efficient cellular energy production and maintenance of cellular health.

Pathways

AKAP1 engages in both the cAMP-dependent and mitochondrial signaling pathways. In the cAMP-dependent pathway AKAP1 interacts with PKA to modulate phosphorylation events that alter cellular responses to external signals. The mitochondrial pathway involves communication with proteins like Drp1 influencing mitochondrial fission and fusion processes. These interactions highlight AKAP1's central role in maintaining cellular energy balance and integrity.

Certain pathways involving AKAP1 link closely with neurodegenerative diseases such as Alzheimer's and cardiac disorders. During neurodegeneration AKAP1-related mitochondrial dysfunction can lead to energy deficits and increased oxidative stress contributing to disease progression. Additionally in cardiac health dysregulation of AKAP1 interactions with PKA can affect heart muscle contractility and lead to cardiac anomalies. Understanding these connections offers insights into potential therapeutic avenues for targeting such diseases.

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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