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AB265782

Human MYOF (Myoferlin) knockout HeLa cell line

Human MYOF (Myoferlin) knockout HeLa cell line

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MYOF KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 10 bp deletion in exon 6. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
2 Images
Western blot - Human MYOF (Myoferlin) knockout HeLa cell line (AB265782)
  • WB

Lab

Western blot - Human MYOF (Myoferlin) knockout HeLa cell line (AB265782)

Lanes 1-4 : Merged signal (red and green). Green - ab178386 observed at 250180 kDa. Red - loading control ab7291 observed at 50 kDa.

ab178386 Anti-Myoferlin antibody [EPR18887] 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°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-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 (ab265782)

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

false

Sanger Sequencing - Human MYOF (Myoferlin) knockout HeLa cell line (AB265782)
  • Sanger seq

Unknown

Sanger Sequencing - Human MYOF (Myoferlin) knockout HeLa cell line (AB265782)

Homozygous : 10 bp deletion in exon 6.

Key facts

細胞タイプ

HeLa

生物種

Human

組織

Cervix

製品の状態

Liquid

form

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

抗生物質耐性

Puromycin 1µg/mL

疾病

Adenocarcinoma

Reactivity data

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製品の詳細

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.

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
Dry Ice
短期保存温度
-196°C
長期保存温度
-196°C

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.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

初回取り扱いガイドライン

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

継代培養ガイドライン
  • 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.
培養培地

DMEM (High Glucose) + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

製品プロトコール

Abcam product promise

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