AB277888

Human SMAD3 knockout A549 cell line

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SMAD3 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift = 100%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

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Lab

Western blot - Human SMAD3 knockout A549 cell line (AB277888)

False colour image of Western blot : Anti-Smad3 antibody [EP568Y] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab40854 was shown to bind specifically to Smad3. A band was observed at 50 kDa in wild-type A549 cell lysates with no signal observed at this size in SMAD3 CRISPR-Cas9 edited cell line ab277888 (CRISPR-Cas9 edited cell lysate None). The band observed in the CRISPR-Cas9 edited lysate lane below 50 kDa is likely to represent a truncated form of Smad3. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and SMAD3 CRISPR-Cas9 edited A549 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-Smad3 antibody [EP568Y] (<a href='/ko/products/primary-antibodies/smad3-antibody-ep568y-ab40854'>ab40854</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

SMAD3 CRISPR-Cas9 edited A549 cell lysate at 20 µg

Secondary

Lanes 1 - 2:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 48 kDa

Observed band size: 50 kDa

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Lab

Western blot - Human SMAD3 knockout A549 cell line (AB277888)

All lanes:

Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (<a href='/ko/products/primary-antibodies/smad3-antibody-epr19686-chip-grade-ab208182'>ab208182</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

SMAD3 CRISPR-Cas9 edited A549 cell lysate at 20 µg

Secondary

Lanes 1 - 2:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 50 kDa

false

주요 정보

세포 유형

A549

Species or organism

Human

조직

Lung

제형

Liquid

form

Mutation 정보

Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift = 100%

질병

Carcinoma

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제품 세부 정보

We will provide viable cells that proliferate on revival.

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.

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": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab277888-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab275463 Human wild-type A549 cell line", "number":"AB277888-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab277888 Human SMAD3 knockout A549 cell line", "number":"AB277888-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab277888-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab277888 Human SMAD3 knockout A549 cell line", "number":"AB277888-CMP01", "productcode":"" } ] } } }

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특성 및 보관 정보

유전자명

SMAD3

유전자 편집 종류

Knockout

유전자 편집 방식

CRISPR technology

품질 관리 - STR analysis

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

배송 시 보관 조건

Dry Ice

적절한 단기 보관 조건

-196°C

적절한 장기 보관 조건

-196°C

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추가 정보

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

Smad3 also known as Mothers against decapentaplegic homolog 3 is a protein that plays a mechanical role in signal transduction. It acts mainly as a transcription factor and gets activated through phosphorylation. The molecular weight of Smad3 is approximately 48 kDa. It is expressed widely across numerous tissues including the cellular nucleus where it executes its function after activation.

Biological function summary

Smad3 acts as a mediator of signal transduction for the TGF-beta (transforming growth factor-beta) superfamily forming a complex with phosphorylated Smad2. This enables it to regulate transcriptional activity influencing cell proliferation differentiation and apoptosis. Smad3 also participates in various cellular processes by interacting with other co-factors and regulatory proteins that aid in fine-tuning its function.

Pathways

Smad3 plays an important role in the TGF-beta signaling pathway where it works closely with Smad4 to propagate the signal. Upon phosphorylation it forms a complex with co-Smad (Smad4) and moves into the nucleus to influence gene expression. Smad3 is also involved in pathways related to oncogenesis and tissue fibrosis indicating its significant role in cellular regulation and response mechanisms.

Smad3 is associated with fibrotic diseases and cancers particularly in tissues such as the liver and lungs. Altered Smad3 signaling contributes to the pathological process occurring in fibrotic disorders often interacting with Smad4 in these abnormalities. Dysregulated Smad3 expression or mutations can also lead to oncogenic transformations highlighting its critical involvement in disease states.

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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.
Do not allow the cell density to exceed 7x10⁴ cells/cm².

배양 배지

F-12K + 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 SMAD3

대체 명칭 보기

Mothers against decapentaplegic homolog 3, MADH3, MAD homolog 3, SMAD 3, Smad3, hMAD-3, SMAD3, SMAD family member 3, Mothers against DPP homolog 3, hSMAD3, Mad3, JV15-2

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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.

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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 SMAD3 knockout A549 cell line

Western blot - Human SMAD3 knockout A549 cell line (AB277888)

Western blot - Human SMAD3 knockout A549 cell line (AB277888)

주요 정보

세포 유형

Species or organism

조직

제형

Mutation 정보

질병

보완 제품

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

제품 세부 정보

제품 구성

특성 및 보관 정보

유전자명

유전자 편집 종류

유전자 편집 방식

품질 관리 - STR analysis

배송 시 보관 조건

적절한 단기 보관 조건

적절한 장기 보관 조건

추가 정보

Biological function summary

Pathways

Cell culture

Biosafety level

Adherent/suspension

Gender

초기 취급 지침

Subculture 가이드라인

배양 배지

동결보존 배지

제품 프로토콜

타겟 정보

대체 명칭 보기

Product promise