AB257103

Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate

Name: Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate (ab257103) | Abcam
Brand: Abcam Limited
SKU: AB257103
Price: 645 USD
Availability: InStock

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NEFM KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.

View Alternative Names

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

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Western blot - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate (AB257103)

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2 : NEFM knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate (20 ug)

ab7794 was shown to specifically react with 160 kD Neurofilament Medium in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266741 (knockout cell lysate ab257103) was used. Wild-type and 160 kD Neurofilament Medium knockout samples were subjected to SDS-PAGE. ab7794 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4^oC at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (<a href='/en-us/products/primary-antibodies/160-kd-neurofilament-medium-antibody-nf-09-neuronal-marker-ab7794'>ab7794</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

NEFM knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-nefm-160-kd-neurofilament-medium-knockout-hek-293t-cell-line-ab266741'>ab266741</a>)

Lane 3:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 102 kDa

Observed band size: 150 kDa

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Sanger Sequencing - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate (AB257103)

Sanger seq

Unknown

Sanger Sequencing - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate (AB257103)

Allele-1 : 1 bp insertion in exon 2

Sanger seq

Unknown

Sanger Sequencing - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate (AB257103)

Allele-2 : Insertion of the selection cassette in exon 2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

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.

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Reactivity data

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

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name

NEFM

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Shipped at conditions

Ambient - Can Ship with Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

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Supplementary information

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

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.

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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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

For licensing inquiries, please contact partnerships@abcam.com

Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate

Western blot - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate (AB257103)

Sanger Sequencing - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate (AB257103)

Sanger Sequencing - Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate (AB257103)

Key facts

Cell type

Species or organism

Tissue

Knockout validation

Mutation description

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Complementary Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Secondary Antibodies

Product promise