ITM2B KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 20 bp insertion in exon 1.
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, 1 bp deletion in exon 1 and 20 bp insertion in exon 1
Recommended products
ITM2B KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 20 bp insertion in exon 1.
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, 1 bp deletion in exon 1 and 20 bp insertion in exon 1
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- ITM2B
- 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 (ab255448). 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
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
ITM2B also known as Integral Membrane Protein 2B or BRI2 is a transmembrane protein weighing approximately 30 kDa. This protein is expressed widely in human tissues showing particularly high levels in the central nervous system and brain. ITM2B functions mechanically by playing a role in protein processing and trafficking owing to its transmembrane nature. With an integral role in cellular functions ITM2B interacts with various cellular components associated with membrane-related activities.
- Biological function summary
ITM2B engages in several vital cellular processes such as regulation of amyloid precursor protein (APP) processing. ITM2B forms part of a complex with other proteins involved in APP cleavage which is significant for maintaining cell homeostasis. It also influences neuronal functions through its effects on synaptic plasticity and could impact cognitive abilities. The protein's presence in brain regions ties ITM2B closely to neural activities where protein interactions are essential for cellular communication.
- Pathways
ITM2B exerts influence on critical processes in amyloidogenic pathways contributing to the modulation of amyloid-beta production. This protein directly interacts with other players like APP and presenilin 1 within this pathway affecting the levels of amyloids and ultimately impacting neural integrity. Additionally ITM2B participates in Wnt signaling pathways linking it to cellular differentiation and proliferation where it modulates interactions with proteins that convey Wnt signals such as beta-catenin.
- Associated diseases and disorders
ITM2B is directly linked to familial British dementia and familial Danish dementia both neurodegenerative conditions. These diseases involve mutations in the ITM2B gene leading to abnormal protein aggregates that disrupt normal brain function. ITM2B's relationship with amyloid-beta and presenilin 1 also connects it to Alzheimer's disease pathogenesis where these proteins play roles in amyloid plaque formation a hallmark of Alzheimer's. Studies focus on targeting ITM2B interactions and processing to develop therapies for these conditions highlighting its potential as a therapeutic target.
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2 product images
Sanger Sequencing - Human ITM2B knockout HeLa cell line (ab266032)
Allele-2: 20 bp insertion in exon 1.
Sanger Sequencing - Human ITM2B knockout HeLa cell line (ab266032)
Allele-1: 1 bp deletion in exon1
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