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AB308483

Mouse Smcr8 knockout RAW 264.7 cell line

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Smcr8 KO cell line available to order. KO validated. Free of charge wild type control available. Homozygote, 1025 bp deletion. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

View Alternative Names

2310076G09Rik, AI642055, D030073L15Rik, RGD1564621, RP23-278I21.6, SMCR8_HUMAN, Smith-Magenis syndrome chromosomal region candidate gene 8 protein

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Sanger Sequencing - Mouse Smcr8 knockout RAW 264.7 cell line (AB308483)
  • Sanger seq

Supplier Data

Sanger Sequencing - Mouse Smcr8 knockout RAW 264.7 cell line (AB308483)

Homozygote, 1025 bp deletion

Key facts

Cell type

RAW 264.7

Species or organism

Mouse

Tissue

Lymphatic

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Homozygote, 1025 bp deletion

Disease

Carcinoma

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Sanger seq": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

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.

Properties and storage information

Gene name
Smcr8
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
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

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.

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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.

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Male

Product protocols

Product promise

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