AB255446

Human VIM (Vimentin) knockout HeLa cell line

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VIM 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: Insertion of the selection cassette in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

別名を表示する

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

3 Images

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Western blot - Human VIM (Vimentin) knockout HeLa cell line (AB255446)

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

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° 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 (ab255446)

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

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Lab

Western blot - Human VIM (Vimentin) knockout HeLa cell line (AB255446)

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° 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 (ab255446)

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 line (AB255446)

Sanger seq

Unknown

Sanger Sequencing - Human VIM (Vimentin) knockout HeLa cell line (AB255446)

Homozygous : Insertion of the selection cassette in exon 2.

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: Insertion of the selection cassette in exon 2

疾病

Adenocarcinoma

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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, 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

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

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取り扱い方法

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.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
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.
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.
4. Incubate the culture at 37°C incubator with 5% CO₂. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x10⁴ cells/cm². Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

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.

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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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ターゲットの情報

See full target information VIM

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Abcam product promise

当社は、高品質な試薬を通じてお客様の研究を力強くサポートすることをお約束いたします。ご使用いただく各段階で、常にお客様をサポートできる体制を整えております。万が一、製品が期待通りに機能しない場合は、「Abcam Product Promise」による当社保証制度に基づき、安心してご利用いただけます。

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'.

For licensing inquiries, please contact partnerships@abcam.com

Human VIM (Vimentin) knockout HeLa cell line