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AB257100

Human FUS (TLS) knockout HEK-293T cell lysate

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FUS 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 deletion in exon2.
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Western blot - Human FUS (TLS) knockout HEK-293T cell lysate (AB257100)
  • WB

Lab

Western blot - Human FUS (TLS) knockout HEK-293T cell lysate (AB257100)

Lane 1 : Wild-type HEK-293T cell lysate (20μg)
Lane 2 : FUS knockout HEK-293T cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab154141 observed at 80 kDa. Red - loading control ab181602 observed at 37 kDa.
ab154141 Anti-TLS/FUS antibody [CL0190] was shown to specifically react with TLS/FUS in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266587 (knockout cell lysate ab257100) was used. Wild-type and TLS/FUS knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154141 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TLS/FUS antibody [CL0190] (<a href='/en-us/products/primary-antibodies/tls-fus-antibody-cl0190-ab154141'>ab154141</a>) at 1/500 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

FUS knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human FUS (TLS) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-fus-tls-knockout-hek-293t-cell-line-ab266587'>ab266587</a>)

Predicted band size: 53 kDa

Observed band size: 80 kDa

false

Sanger Sequencing - Human FUS (TLS) knockout HEK-293T cell lysate (AB257100)
  • Sanger seq

Unknown

Sanger Sequencing - Human FUS (TLS) knockout HEK-293T cell lysate (AB257100)

Homozygous : 1 bp deletion in exon2

Western blot - Human FUS (TLS) knockout HEK-293T cell lysate (AB257100)
  • WB

Lab

Western blot - Human FUS (TLS) knockout HEK-293T cell lysate (AB257100)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)
Lane 2 :  FUS knockout HEK-293T cell lysate (20µg)
Lanes 1- 2 : Merged signal (red and green). Green - ab124923 observed at 75 kDa. Red - loading control ab8245 observed at 37 kDa.
ab124923 Anti-TLS/FUS antibody [EPR5812] was shown to specifically react with TLS/FUS in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266587 (knockout cell lysate ab257100) was used. Wild-type and TLS/FUS knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab124923 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TLS/FUS antibody [EPR5812] (<a href='/en-us/products/primary-antibodies/tls-fus-antibody-epr5812-ab124923'>ab124923</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human FUS (TLS) knockout HEK-293T cell lysate (ab257100) at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 75 kDa,37 kDa

false

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 deletion in exon2.

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Complementary Products

Primary Antibodies

AB124923

Anti-TLS/FUS antibody [EPR5812]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [EPR5812] (AB124923)

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Proteins & Peptides

AB259412

Recombinant human VEGF 165A protein (Active)

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AB288799

Recombinant Human VCAM1 protein (Active)

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Biological Activity - Recombinant Human VCAM1 protein (Active) (AB288799)

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Proteins & Peptides

AB52101

Recombinant human Cleaved Caspase-3 protein (Active)

Western blot - Recombinant human Cleaved Caspase-3 protein (Active) (AB52101)

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Reactivity data

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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 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.

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What's included?

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Properties and storage information

Gene name
FUS
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.

TLS/FUS also known as FUS and the FUS protein is an RNA-binding protein involved in various cellular processes. It has a molecular weight of approximately 53 kDa. Expressed extensively in the nucleus FUS/TLS relocates to the cytoplasm under stress conditions. It plays mechanical roles in transcription regulation RNA splicing and mRNA transport. Scientists sometimes refer to FUS as the 'fused in sarcoma' protein because of its involvement in gene fusion events.
Biological function summary

The FUS protein aids in the maintenance and stabilization of mRNA molecules and participates in the formation of stress granules distinct cytoplasmic aggregates of proteins and RNAs. It forms part of a large protein complex alongside other RNA-binding proteins. FUS stabilizes pre-mRNA structures which affects protein expression patterns essential for cell survival and differentiation. Its ability to bind to RNA and DNA indicates its fundamental role in genomic stability.

Pathways

The functions of FUS help regulate RNA metabolism-related pathways and cellular stress response pathways. FUS interacts with proteins like TAF15 and EWSR1 within these pathways showing a complex network of interactions that contribute to RNA maturation and stress granule dynamic formation. The activity of FUS in these pathways ultimately affects cellular homeostasis gene expression modulation and response to cellular stress.

FUS protein mutations or mislocalizations relate to neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). These conditions are characterized by the accumulation of FUS or FUS-related aggregates in the cytoplasm leading to neuronal cell death. FUS is also connected to other RNA-binding proteins like TDP-43 which also mislocalizes and forms aggregates in these disorders highlighting the importance of protein homeostasis and normal RNA metabolism in the prevention of these diseases.
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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

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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'.

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