AB257547

Human MYOF (Myoferlin) knockout HeLa cell lysate

Name: Human MYOF (Myoferlin) knockout HeLa cell lysate (ab257547) | Abcam
Brand: Abcam Limited
SKU: AB257547
Price: 114000 JPY
Availability: InStock

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MYOF KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 10 bp deletion in exon 6.

2 Images

Lab

Western blot - Human MYOF (Myoferlin) knockout HeLa cell lysate (AB257547)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : MYOF knockout HeLa cell lysate (20 ug)
Lane 3 : MDA-MB-231 cell lysate (20 ug)
Lane 4 : SK-BR-3 cell lysate (20 ug)

ab178386 was shown to specifically react with Myoferlin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265782 (knockout cell lysate ab257547) was used. Wild-type and Myoferlin knockout samples were subjected to SDS-PAGE. ab178386 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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^® 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-Myoferlin antibody [EPR18887] (<a href='/products/primary-antibodies/myoferlin-antibody-epr18887-ab178386'>ab178386</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MYOF knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MYOF (Myoferlin) knockout HeLa cell line (<a href='/products/cell-lines/human-myof-myoferlin-knockout-hela-cell-line-ab265782'>ab265782</a>)

Lane 3:

MDA-MB-231 cell lysate at 20 µg

Lane 4:

SK-BR-3 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 235 kDa

Observed band size: 180 kDa,250 kDa

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Sanger Sequencing - Human MYOF (Myoferlin) knockout HeLa cell lysate (AB257547)

Sanger seq

Unknown

Sanger Sequencing - Human MYOF (Myoferlin) knockout HeLa cell lysate (AB257547)

Homozygous : 10 bp deletion in exon 6

Key facts

細胞タイプ

HeLa

生物種

Human

組織

Cervix

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, Homozygous: 10 bp deletion in exon 6.

疾病

Adenocarcinoma

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Reactivity data

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製品の詳細

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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製品内容

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出荷温度及び保存条件

遺伝子名

MYOF

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing, Western blot

接合型

Homozygous

出荷温度

Ambient - Can Ship with Ice

短期保存温度

-20°C

長期保存温度

-20°C

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補足情報

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

Myoferlin alternatively known as MYOF is a member of the ferlin family proteins and plays a mechanical role in membrane repair and fusion. It has a molecular mass of approximately 230 kDa. Myoferlin expresses mainly in muscle and endothelial tissues. Its function is critical during muscle development and regeneration as it participates in the fusion of myoblasts into mature myotubes. Also Myoferlin is involved in the repair of damaged sarcolemmal membranes in muscle cells.

Biological function summary

The protein supports membrane dynamics and stabilization. Myoferlin is not part of a large complex but interacts with other proteins at the cellular membrane. It regulates cellular processes such as proliferation differentiation and endocytosis. In muscle cells it assists in the organization and function of the cytoskeleton maintaining proper cellular architecture and signaling.

Pathways

Myoferlin engages in critical roles in pathways related to cell membrane repair and trafficking. It associates with pathways like the myogenesis pathway and integrin signaling. In these pathways it interacts with other proteins such as dysferlin and caveolin-3 which facilitate membrane repair and receptor localization respectively. Myoferlin ensures efficient cellular response to membrane damage maintaining tissue integrity.

Myoferlin is connected to muscular dystrophies and cancer. It correlates with the progression of muscular disorder due to its deficiency in membrane repair mechanisms. Additionally in cancer its expression levels relate to tumor invasion and metastasis involving interactions with proteins like E-cadherin and MMP-9. Investigating Myoferlin's role can aid in understanding its potential as a therapeutic target in 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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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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Abcam product promise

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Human MYOF (Myoferlin) knockout HeLa cell lysate