AB286576

Human CASP8 knockout HCT116 cell line

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CASP8 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

대체 명칭 보기

FADD-homologous ICE/ced-3-like protease, MORT1-associated ced-3 homolog, CASP-8, FLICE, Apoptotic cysteine protease, CAP4, FADD-like ICE, MACH, CASP8, Caspase-8, ICE-like apoptotic protease 5, MCH5, Apoptotic protease Mch-5

5 이미지

Lab

Western blot - Human CASP8 knockout HCT116 cell line (AB286576)

Western blot : Anti-CASP8 antibody [E7] (ab32397) staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab32397 was shown to bind specifically to CASP8. A band was observed at 55 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CASP8 knockout cell line. To generate this image, wild-type and CASP8 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Caspase-8 antibody [E7] (<a href='/ko/products/primary-antibodies/caspase-8-antibody-e7-ab32397'>ab32397</a>) at 1/500 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

CASP8 knockout HCT 116 cell lysate at 20 µg

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

CASP8 knockout HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

false

Lab

Western blot - Human CASP8 knockout HCT116 cell line (AB286576)

Western blot : Rabbit Monoclonal[E6] to Caspase-8 ab32125 staining at 1/3000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 55 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in CASP8 knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^{^®} 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Caspase-8 antibody [E6] (<a href='/ko/products/primary-antibodies/caspase-8-antibody-e6-ab32125'>ab32125</a>) at 1/3000 dilution

Lane 1:

Wild-type HCT 116 at 20 µg

Lane 2:

Western blot - Human CASP8 knockout HCT116 cell line (ab286576) at 20 µg

Lane 3:

Wild-type HeLa at 20 µg

Lane 4:

CASP8 knockout HeLa at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

false

Lab

Next Generation Sequencing - Human CASP8 knockout HCT116 cell line (AB286576)

118 bp deletion after Pro30

Lab

Western blot - Human CASP8 knockout HCT116 cell line (AB286576)

Western blot : Anti-Caspase-8 antibody [E6] ab32125 staining at 1/3000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 60 kDa in Wild-type A549 cell lysates with no signal observed at this size in CASP8 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Caspase-8 antibody [E6] (<a href='/ko/products/primary-antibodies/caspase-8-antibody-e6-ab32125'>ab32125</a>) at 1/3000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

Western blot - Human CASP8 knockout A549 cell line (<a href='/ko/products/cell-lines/human-casp8-knockout-a549-cell-line-ab286757'>ab286757</a>) at 20 µg

Lane 3:

Western blot - Human wild-type HCT116 cell line (ab288559) at 20 µg

Lane 4:

Western blot - Human CASP8 knockout HCT116 cell line (ab286576) at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 60 kDa,37 kDa

Observed band size: 60 kDa

false

Lab

Western blot - Human CASP8 knockout HCT116 cell line (AB286576)

Western blot : Anti-Caspase-8 antibody [E7] ab32397 staining at 1/500 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 60 kDa in Wild-type A549 cell lysates with no signal observed at this size in CASP8 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Caspase-8 antibody [E7] (<a href='/ko/products/primary-antibodies/caspase-8-antibody-e7-ab32397'>ab32397</a>) at 1/500 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

Western blot - Human CASP8 knockout A549 cell line (<a href='/ko/products/cell-lines/human-casp8-knockout-a549-cell-line-ab286757'>ab286757</a>) at 20 µg

Lane 3:

Western blot - Human wild-type HCT116 cell line (ab288559) at 20 µg

Lane 4:

Western blot - Human CASP8 knockout HCT116 cell line (ab286576) at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 60 kDa

Observed band size: 60 kDa,37 kDa

false

주요 정보

세포 유형

HCT116

Species or organism

Human

조직

Colon

제형

Liquid

form

Knockout 검증

Next Generation Sequencing,Western blot

질병

Carcinoma

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Reactivity 정보

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" } } }

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제품 세부 정보

Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.

Recommended control: Human wild-type HCT116 cell line (ab288559). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

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.

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제품 구성

{ "values": { "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab286576-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"Human CASP8 knockout HCT116 cell line kit component", "number":"AB286576-CMP01" } ] }, "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab286576-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"Human CASP8 knockout HCT116 cell line kit component", "number":"AB286576-CMP01", "productcode":"" }, { "size":"1 x 1000000 Cells/vial", "name":"Human wild-type HCT116 cell line kit component", "number":"AB286576-CMP02", "productcode":"" } ] } } }

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특성 및 보관 정보

유전자명

CASP8

유전자 편집 종류

Knockout

유전자 편집 방식

CRISPR technology

Knockout 검증

Next Generation Sequencing, Western blot

배송 시 보관 조건

Dry Ice

적절한 단기 보관 조건

-196°C

적절한 장기 보관 조건

-196°C

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품질 관리

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

초기 취급 지침

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.

Subculture 가이드라인

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.

배양 배지

McCoY5a + 10% FBS

동결보존 배지

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

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제품 프로토콜

Visit the General protocols
Visit the Troubleshooting

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타겟 정보

See full target information CASP8

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Product promise

저희는 고품질 시약으로 고객님의 연구를 서포트하고자 최선을 다하고 있으며, 모든 단계에서 함께 하겠습니다. 만약 제품이 기대한 성능을 보이지 않을 경우에도 Product Promise로 보호받으실 수 있습니다.

자세한 내용은 이용 약관을 확인해 주세요.

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 CASP8 knockout HCT116 cell line

Western blot - Human CASP8 knockout HCT116 cell line (AB286576)

Western blot - Human CASP8 knockout HCT116 cell line (AB286576)

Next Generation Sequencing - Human CASP8 knockout HCT116 cell line (AB286576)

Western blot - Human CASP8 knockout HCT116 cell line (AB286576)

Western blot - Human CASP8 knockout HCT116 cell line (AB286576)

주요 정보

세포 유형

Species or organism

조직

제형

Knockout 검증

질병

보완 제품

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity 정보

제품 세부 정보

제품 구성

특성 및 보관 정보

유전자명

유전자 편집 종류

유전자 편집 방식

Knockout 검증

배송 시 보관 조건

적절한 단기 보관 조건

적절한 장기 보관 조건

품질 관리

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

초기 취급 지침

Subculture 가이드라인

배양 배지

동결보존 배지

제품 프로토콜

타겟 정보

Product promise