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NEFM 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 insertion in exon 2 and Insertion of the selection cassette in exon 2.

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Images

Western blot - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell line (AB266741), expandable thumbnail
  • Cell Culture - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell line (AB266741), expandable thumbnail
  • Sanger Sequencing - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell line (AB266741), expandable thumbnail
  • Sanger Sequencing - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell line (AB266741), expandable thumbnail

Key facts

Cell type
HEK-293T
Species or organism
Human
Tissue
Kidney
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2

Alternative names

150kDa medium, 160 kDa neurofilament protein, NEF3, NEFM, NF160, NFM_HUMAN, Neurofilament 3, Neurofilament medium polypeptide, Neurofilament triplet M protein

Recommended products

NEFM 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 insertion in exon 2 and Insertion of the selection cassette in exon 2.

Key facts

Cell type
HEK-293T
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2
Concentration
Loading...

Properties

Gene name
NEFM
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot

Quality control

STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level
EU: 2 US: 2
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

Recommended control: Human wild-type HEK293T cell line (ab255449). 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, 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

The 160 kD Neurofilament Medium also known as NEFM NF-M or NEF-M belongs to the family of neurofilament proteins. This protein is an important structural component of neurons particularly in the axonal cytoskeleton. Neurofilament Medium has a molecular mass of around 160 kilodaltons (kD). It shows high levels of expression in neuronal tissues contributing to the maintenance of axonal caliber. Commonly research recognizes it alongside other neurofilament proteins emphasizing its role in providing structural integrity to neurons.

Biological function summary

Neurofilament Medium participates in the formation of a stable network of neurofilaments within neurons. Acting as a part of the intermediate filament protein family it forms a complex with other neurofilaments such as NF-L and NF-H. This complex supports neuron structure and plays an important role in axonal transport. The heteropolymeric nature of neurofilaments contributes to their mechanical stability facilitating essential neuronal functions by aligning neurofilaments along the axon.

Pathways

Neurofilament Medium integrates into the cytoskeletal arrangement pathways that govern axonal transport and stability. It plays a significant role in the neuronal transport pathway associating with other proteins such as dynein and kinesin responsible for the motor functions along axons. Also it relates to the MAP kinase pathway where phosphorylation events modulate its assembly and disassembly dynamics in response to cellular signals.

Associated diseases and disorders

Neurofilament Medium closely relates to neurodegenerative conditions such as amyotrophic lateral sclerosis (ALS) and Alzheimer's disease. Aberrant phosphorylation or aggregation of NEFM can disrupt neuronal function leading to pathology. In ALS the abnormal accumulation of neurofilaments correlates with motor neuron degeneration. In Alzheimer's its interaction with tau protein poses significant interest as altered states of either could exacerbate neurofibrillary tangles worsening cognitive decline.

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Product protocols

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

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