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SHMT2 KO cell line available to order. Free of charge wild type control provided.

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Key facts

Cell type
A549
Species or organism
Human
Tissue
Lung
Form
Liquid

Alternative names

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SHMT2 KO cell line available to order. Free of charge wild type control provided.

Key facts

Cell type
A549
Form
Liquid
Disease
Carcinoma
Concentration
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Properties

Gene name
SHMT2
Gene editing type
Knockout
Gene editing method
CRISPR technology

Cell culture

Biosafety level
EU: 1 US: 1
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 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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
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 A549 cell line (ab288558). 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.

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

Serine hydroxymethyltransferase 2 (SHMT2) also known as SHMT is an enzyme that plays a role in the mitochondrial compartment. It has a molecular mass of approximately 53 kDa. SHMT2 is highly expressed in tissues with elevated rates of oxidative metabolism such as the liver and brain. The enzyme catalyzes the reversible conversion of serine to glycine with the concomitant production of 510-methylenetetrahydrofolate a critical molecule for DNA synthesis and repair.

Biological function summary

The enzyme facilitates essential reactions in mitochondrial nucleotide synthesis. SHMT2 operates within a tetrameric complex that provides one-carbon units for cellular biosynthetic processes. The activity of SHMT2 links mitochondrial serine utilization directly to the folate cycle which is essential for the synthesis of purines and thymidylate. The efficient production of these nucleotides is important for rapidly dividing cells supporting their proliferation and maintenance.

Pathways

SHMT2 takes part in the folate and methionine metabolism pathways. It interacts with proteins like methylenetetrahydrofolate dehydrogenase and thymidylate synthase which are essential for one-carbon metabolism. These interactions integrate SHMT2 activity into a larger network that balances the supply of nucleotides according to the cellular demand impacting vital pathways like DNA replication and repair.

Associated diseases and disorders

Elevated SHMT2 expression has been linked to cancer where it supports the rapid proliferation of tumor cells by providing necessary nucleotides. Its role in the mitochondrial folate pathway is also significant in neurodegenerative disorders. SHMT2 in conjunction with other proteins such as methionine synthase may play a role in the pathology of diseases like Alzheimer's by influencing homocysteine levels and oxidative stress in brain tissues.

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.

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    For this product, it's our understanding that no specific protocols are required. You can:

    Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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