ADCY3 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1
Alternative names
AC-III, ADCY3_HUMAN, ATP pyrophosphate-lyase, ATP pyrophosphate-lyase 3, Adenylate cyclase, Adenylate cyclase type 3, Adenylate cyclase type III, Adenylyl cyclase 3, adenylate cyclase 3, olfactive type
Recommended products
ADCY3 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- ADCY3
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- Zygosity
- Homozygous
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 short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The protein AC3 also known as 'Adcy3' functions as an adenylate cyclase enzyme. This enzyme plays a significant mechanical role by converting ATP (adenosine triphosphate) to cyclic AMP (cAMP) which acts as a secondary messenger in various signaling pathways. It is also referred to as '1a12' in some studies. Weighing approximately 137 kDa AC3 is mainly expressed in the olfactory epithelium where it contributes to odor detection the brain and other tissues at lower levels. This protein operates at the cell membrane where it interacts with G proteins to mediate signal transduction processes.
- Biological function summary
Adenylate cyclase 3 (AC3) serves as an essential component in sensory signal transduction and is part of the adenylyl cyclase complex. It plays an important role in the conversion of external stimuli into intracellular signals important for processes like olfaction and hormonal response. Within neurons AC3 influences synaptic plasticity and learning by regulating cAMP levels. This modulation affects various neural activities and responses making AC3 important for central nervous system functions.
- Pathways
AC3 is integrally involved in the G-protein coupled receptor (GPCR) signaling pathways. These pathways involve the activation of AC3 by G-proteins leading to enhanced production of cAMP which subsequently activates protein kinase A (PKA). AC3 collaborates with other adenylate cyclases and G-protein subunits such as Gα and Gβγ within these pathways. This intricate signaling cascade influences cellular responses to hormones and neurotransmitters and alters gene expression through cAMP-mediated pathways.
- Associated diseases and disorders
AC3 has associations with conditions like obesity and depression. Abnormalities in AC3 function or expression can lead to impaired cAMP signaling disrupting normal metabolic and neurological processes. Research indicates that AC3 interacts with the melanocortin-4 receptor (MC4R) in regulating energy homeostasis and body weight linking it to obesity. In depression altered AC3 levels impact neurotransmitter signaling contributing to mood disorders. Understanding AC3's role in these conditions may offer therapeutic insights into disease management.
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2 product images
Sanger Sequencing - Human ADCY3 (AC3) knockout HeLa cell line (ab264709)
Homozygous: 1 bp deletion in exon 1.
Immunocytochemistry/ Immunofluorescence - Human ADCY3 (AC3) knockout HeLa cell line (ab264709)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized ADCY3 KO HeLa (ADCY3 knockout human cervical adenocarcinoma epithelial cell), ab264709 cells labelling AC3 with Anti-AC3 antibody [EPR28777-12] ab323453 at 1/2000 (0.263 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ 1000 2ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in wildtype HeLa cells (shown in green), showing no staining in ADCY3 knockout HeLa cells. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ 200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ 1000 2ug/ml dilution.
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