Smcr8 KO cell line available to order. Free of charge wild type control provided. Homozygote, 1025 bp deletion.
Images
Key facts
- Cell type
- RAW 264.7
- Species or organism
- Mouse
- Tissue
- Lymphatic
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Homozygote, 1025 bp deletion
Alternative names
2310076G09Rik, AI642055, D030073L15Rik, RGD1564621, RP23-278I21.6, SMCR8_HUMAN, Smith-Magenis syndrome chromosomal region candidate gene 8 protein
Smcr8 KO cell line available to order. Free of charge wild type control provided. Homozygote, 1025 bp deletion.
Key facts
- Cell type
- RAW 264.7
- Species or organism
- Mouse
- Tissue
- Lymphatic
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Homozygote, 1025 bp deletion
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- Smcr8
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Male
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 for bath 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 all remaining cells into an ultra-low attachment 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.
5. Once confluent passage into an appropriate flask at a density of 2x105 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
- Grow in ultra-low attachment flasks.
- Do not wash or use dissociation reagent. Cells are semi-adherent. Tap flask sharply to remove adhered cells.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x105 cells/mL is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- DMEM + 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 duration
- A few days
- Appropriate short-term storage conditions
- -80°C
- Appropriate long-term storage conditions
- -196°C
Notes
Recommended control: Human wild-type RAW 264.7 cell line (ab275474). 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 + 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 an ultra-low attachment 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.
5. Once confluent passage into an appropriate flask at a density of 2x105 cells / mL. 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:
- Grow in ultra-low attachment flasks.
- Do not wash or use dissociation reagent. Cells are semi-adherent. Tap flask sharply to remove adhered cells.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x10e5 cells/mL is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
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
SMCR8 also known as Smith-Magenis Syndrome Chromosome Region 8 is a protein with a molecular weight of approximately 105 kDa. This protein is expressed in multiple tissues with significant presence in the brain kidney and liver. SMCR8 functions mechanically as a guanine nucleotide exchange factor and it plays a vital role in cellular homeostasis and autophagy.
- Biological function summary
SMCR8 is involved in regulating macroautophagy a critical process for cellular maintenance by forming a complex with the C9orf72 protein. This complex ensures proper lysosomal positioning and function impacting the cell's ability to remove damaged organelles and proteins. SMCR8’s activity is essential for maintaining normal lysosomal dynamics and preventing the accumulation of toxic substances within the cell.
- Pathways
SMCR8 participates in the autophagy and lysosome pathways. It interacts with several other proteins including UVRAG and Beclin1 which are important for autophagosome formation and maturation. SMCR8 and its associated pathways are important in coordinating proper cellular degradation and recycling processes ensuring cellular stability and function.
- Associated diseases and disorders
SMCR8 has been connected to neurodegenerative conditions such as amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Mutations or dysfunction in SMCR8 can interfere with autophagy contributing to the pathogenesis of these disorders. The interaction between SMCR8 and C9orf72 has been particularly highlighted in relation to ALS as disruptions in this interaction could aggravate disease progression.
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1 product image
Sanger Sequencing - Mouse Smcr8 knockout RAW 264.7 cell line (ab308483)
Homozygote, 1025 bp deletion
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