BACH1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 38 bp deletion in exon 2 and 43 bp deletion in exon 2 and 8 bp deletion in exon 2.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 38 bp deletion in exon 2 and 43 bp deletion in exon 2 and 8 bp deletion in exon 2
Recommended products
BACH1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 38 bp deletion in exon 2 and 43 bp deletion in exon 2 and 8 bp deletion in exon 2.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 38 bp deletion in exon 2 and 43 bp deletion in exon 2 and 8 bp deletion in exon 2
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- BACH1
- 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 short-term storage conditions
- -196°C
- 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 HeLa cell line (ab271142). 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 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 BACH1.3 gene encodes a transcription regulator protein also known as BTB and CNC homology 1 basic leucine zipper transcription factor 1. The protein weighs approximately 105 kDa and you can find it in various tissues including the liver lungs and kidneys. It binds to the antioxidant response elements (ARE) in DNA regulating the transcription of genes involved in oxidative stress response. BACH1.3 functions by repressing gene activation through heterodimerization with small Maf proteins which helps maintain cellular redox balance.
- Biological function summary
BACH1.3 modulates gene expression by forming complexes with small Maf proteins which are necessary for its regulatory activity. This protein plays a role in cellular oxidative stress management and heme metabolism. Through its function in stress response regulation BACH1.3 influences the expression of several genes such as heme oxygenase-1 (HO-1) important for detoxifying reactive oxygen species. By controlling these pathways BACH1.3 supports cell protection and adaptation to environmental oxidative stress.
- Pathways
BACH1.3 participates in the oxidative stress response and heme degradation pathways. It partners with the KEAP1-NRF2 pathway where it acts as a negative regulator of NRF2 target genes by competing with NRF2 for binding to ARE sequences. This action impacts the transcription of antioxidant genes. BACH1.3 indirectly affects the expression of proteins like HO-1 and NAD(P)H:quinone oxidoreductase 1 (NQO1) moderating their protective effects in the cell.
- Associated diseases and disorders
Altered BACH1.3 function associates with cancer and cardiovascular diseases. Its involvement in improper oxidative stress response can lead to tumor progression and resistance to therapeutic agents in some cancers. Additionally in cardiovascular diseases BACH1.3 connects to abnormal arterial remodeling due to its regulation of genes involved in oxidative stress. This target interacts with proteins like KEAP1 and NRF2 modifying disease outcomes through changes in redox-sensitive signaling and gene expression.
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3 product images
Sanger Sequencing - Human BACH1 (BACH1.3) knockout HeLa cell line (ab265322)
Allele-1: 38 bp deletion in exon 2.
Sanger Sequencing - Human BACH1 (BACH1.3) knockout HeLa cell line (ab265322)
Allele-2: 8 bp deletion in exon 2.
Sanger Sequencing - Human BACH1 (BACH1.3) knockout HeLa cell line (ab265322)
Allele-3: 43 bp deletion in exon 2.
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