AB263775

Human VIM (Vimentin) knockout HeLa cell lysate

Name: Human VIM (Vimentin) knockout HeLa cell lysate (ab263775) | Abcam
Brand: Abcam Limited
SKU: AB263775
Price: 114000 JPY
Availability: InStock

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VIM KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon2.

別名を表示する

CTRCT30, Epididymis luminal protein 113, FLJ36605, HEL113, VIME_HUMAN, Vimentin

3 Images

Unknown

Western blot - Human VIM (Vimentin) knockout HeLa cell lysate (AB263775)

This image was generated using a previous batch manufactured using the hybridoma production method.

Lane 1 : U20S cell lysate (20 μg)

Lane 2 : Hu tonsil cell lysate (20 μg)

Lane 3 : Wild-type HeLa cell lysate (20 μg)

Lane 4 : VIM knockout HeLa cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab16700 observed at 53 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab16700 was shown to react with Vimentin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255446 (knockout cell lysate ab263775) was used. Wild-type and Vimentin knockout samples were subjected to SDS-PAGE. ab16700 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 100 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.

Lanes 1 - 4:

Western blot - Anti-Vimentin antibody [SP20], prediluted (<a href='/products/primary-antibodies/vimentin-antibody-sp20-prediluted-ab27608'>ab27608</a>) at 1/100 dilution

Lanes 1 - 4:

Western blot - Anti-Vimentin antibody [SP20] (<a href='/products/primary-antibodies/vimentin-antibody-sp20-ab16700'>ab16700</a>) at 1/100 dilution

Lane 1:

U20S cell lysate at 20 µg

Lane 2:

Human tonsil cell lysate at 20 µg

Lane 2:

Western blot - Human VIM (Vimentin) knockout HeLa cell line (<a href='/products/cell-lines/human-vim-vimentin-knockout-hela-cell-line-ab255446'>ab255446</a>)

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

VIM knockout HeLa 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 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 37 kDa,53 kDa

false

Lab

Western blot - Human VIM (Vimentin) knockout HeLa cell lysate (AB263775)

Lane 1 : U20S cell lysate (20 μg)

Lane 2 : Hu tonsil cell lysate (20 μg)

Lane 3 : Wild-type HeLa cell lysate (20 μg)

Lane 4 : VIM knockout HeLa cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab8069 observed at 53 kDa. Red - loading control, ab181602 observed at 37 kDa.

ab8069 was shown to react with Vimentin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255446 (knockout cell lysate ab263775) was used. Wild-type and Vimentin knockout samples were subjected to SDS-PAGE. ab8069 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 μg/ml 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-Vimentin antibody [V9] - Cytoskeleton Marker (<a href='/products/primary-antibodies/vimentin-antibody-v9-cytoskeleton-marker-ab8069'>ab8069</a>) at 1 µg/mL

Lane 1:

U-2 OS (Human bone osteosarcoma epithelial cell line) cell lysate at 20 µg

Lane 2:

Human tonsil cell lysate at 20 µg

Lane 2:

Western blot - Human VIM (Vimentin) knockout HeLa cell line (<a href='/products/cell-lines/human-vim-vimentin-knockout-hela-cell-line-ab255446'>ab255446</a>)

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

VIM knockout HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 50 kDa,53 kDa

false

Sanger Sequencing - Human VIM (Vimentin) knockout HeLa cell lysate (AB263775)

Sanger seq

Unknown

Sanger Sequencing - Human VIM (Vimentin) knockout HeLa cell lysate (AB263775)

Homozygous : Insertion of the selection cassette in exon2

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, Insertion of the selection cassette in exon2.

疾病

Adenocarcinoma

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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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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, 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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製品内容

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出荷温度及び保存条件

遺伝子名

VIM

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

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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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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保証に関する詳細については利用規約をご確認ください。

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

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Human VIM (Vimentin) knockout HeLa cell lysate