PA2G4 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1.
Images
Key facts
- Cell type
HEK-293T
- Species or organism
Human
- Tissue
Kidney
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1
Recommended products
PA2G4 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1.
Key facts
- Cell type
HEK-293T
- Species or organism
Human
- Tissue
Kidney
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1
- Concentration
Properties
- Gene name
PA2G4
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
Cell culture
- Biosafety level
EU: 2 US: 2
- Viability
~ 80%
- 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
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type HEK293T cell line (Human wild-type HEK-293T cell line ab255449). 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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM (High Glucose) + 10% FBS
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 2x10E4 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 2x10E4 cells/cm2 is recommended.
• Cells should be passaged when they have achieved 80-90% confluence.
• Do not allow the cell density to exceed 7x10E4 cells/cm2.
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
The EBP1 protein also known as ErbB3-binding protein 1 plays an important role in cellular mechanisms. With a mass of about 39 kDa EBP1 is widely expressed in various tissues including the heart liver and brain. The protein acts as an RNA-binding protein and it has demonstrated influence in the regulation of ribosomal RNA synthesis and processing. EBP1 also interacts with multiple other proteins which aid in its regulatory roles in the cells.
- Biological function summary
EBP1 functions as a part of the cellular machinery involved in growth and differentiation. It associates with the eukaryotic translation initiation factor 4G (eIF4G) as part of a complex which modulates the translation of certain messenger RNAs. Furthermore EBP1 can function as a transcriptional co-repressor in the nucleus where it influences gene expression by interacting with DNA. Its activity in these roles highlights its important function in cell cycle regulation and cellular proliferation.
- Pathways
Many signaling mechanisms engage EBP1. The PI3K/AKT pathway sees participation from EBP1 where it downstream affects cell survival and growth. EBP1 also interacts with the NF-κB pathway contributing to the regulation of immune and stress responses. In these pathways interactions with key proteins like AKT and NF-κB p65 emphasize its role in integrating signaling cues for responses to environmental stimuli.
- Associated diseases and disorders
EBP1 has shown associations with cancer and neurodegenerative diseases. Its connection with cancer is evident as EBP1 influences cell proliferation and can affect tumor suppressor pathways sometimes interacting with proteins like HER2. In neurodegenerative diseases deregulation of EBP1 function has been seen in conditions such as Alzheimer's potentially interacting with proteins involved in tau phosphorylation. Both examples demonstrate the biological significance of EBP1 in maintaining normal physiological function and its contributions to pathology when dysregulated.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
2 product images
Sanger Sequencing - Human PA2G4 (EBP1) knockout HEK-293T cell line (ab266254)
Allele-1: 1 bp insertion in exon 1
Sanger Sequencing - Human PA2G4 (EBP1) knockout HEK-293T cell line (ab266254)
Allele-2: Insertion of the selection cassette in exon 1.
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