AB256848

Human BBC3 (PUMA) knockout HeLa cell lysate

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BBC3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon2 and 1 bp insertion in exon2 and 22 bp deletion in exon2.

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PUMA, JFY-1, BBC3, p53 up-regulated modulator of apoptosis

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Western blot - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)

Lane 1 : Wild-type HeLa cell lysate 20 ug
Lane 2 : BBC3 knockout HeLa cell lysate 20 ug
Lane 3 : A549 cell lysate 20 ug
Lane 4 : PC-3 cell lysate 20 ug
Lanes 1 - 4 : Merged signal (red and green). Green - anti-Puma antibody observed at 25 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
anti-Puma antibody was shown to react with Puma in wild-type HeLa cells in Western blot with loss of signal observed in BBC3 knockout cell line ab261832 (BBC3 knockout cell lysate ab256848). Wild-type HeLa and BBC3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with anti-Puma antibody and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ^°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Anti-Puma antibody at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

BBC3 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human BBC3 (PUMA) knockout HeLa cell line (<a href='/ko/products/cell-lines/human-bbc3-puma-knockout-hela-cell-line-ab261832'>ab261832</a>)

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

PC-3 cell lysate at 20 µg

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Sanger Sequencing - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)

Sanger seq

Unknown

Sanger Sequencing - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)

Allele-1 : 22 bp deletion in exon2

Sanger seq

Unknown

Sanger Sequencing - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)

Allele-2 : 10 bp deletion in exon2

Sanger seq

Unknown

Sanger Sequencing - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)

Allele-3 : 1 bp insertion in exon2

주요 정보

세포 유형

HeLa

Species or organism

Human

조직

Cervix

Knockout 검증

Sanger Sequencing,Western blot

Mutation 정보

Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon2 and 1 bp insertion in exon2 and 22 bp deletion in exon2.

질병

Adenocarcinoma

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Reactivity 정보

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

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제품 세부 정보

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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제품 구성

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab256848-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB256848-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human BBC3 knockout HeLa cell lysate", "number":"AB256848-CMP01", "productcode":"" } ] } } }

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특성 및 보관 정보

유전자명

BBC3

유전자 편집 종류

Knockout

유전자 편집 방식

CRISPR technology

Knockout 검증

Sanger Sequencing, Western blot

배송 시 보관 조건

Ambient - Can Ship with Ice

적절한 단기 보관 조건

-20°C

적절한 장기 보관 조건

-20°C

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추가 정보

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

PUMA also known as BBC3 (Bcl-2-binding component-3) is a pro-apoptotic protein that plays important mechanical roles in promoting apoptosis. It has a mass of approximately 23 kDa. PUMA is expressed widely in various tissues including the lung liver and heart. The protein works mechanically by interacting with anti-apoptotic Bcl-2 family members releasing pro-apoptotic factors like Bax and Bak which engage mitochondria to trigger cell death.

Biological function summary

PUMA functions as a pivotal mediator of apoptosis in response to diverse stimuli such as DNA damage and oncogenic stress. As part of the Bcl-2 family PUMA does not typically form a complex with other proteins but functions directly by binding and neutralizing anti-apoptotic Bcl-2 family proteins. This action results in the release of caspase-activating factors from mitochondria essential for programmed cell death.

Pathways

Several cellular pathways incorporate PUMA to regulate apoptosis. PUMA is notably included in the p53 pathway where p53 transcriptionally activates PUMA following cellular stress or DNA damage. The presence of related proteins like Bax and Bim in the mitochondrial apoptotic pathway links these signals to the mitochondrial death machinery facilitating apoptosis.

PUMA plays significant roles in cancer and neurodegenerative conditions. PUMA overexpression induces excessive cell death relevant in the pathology of neurodegenerative diseases. In cancer PUMA's interactions with proteins like p53 highlight its involvement in tumor suppression suggesting that dysregulation of PUMA expression can contribute to tumorigenesis when anti-apoptotic signals predominate potentially leading to chemoresistance.

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품질 관리

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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제품 프로토콜

Visit the General protocols
Visit the Troubleshooting

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Product promise

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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 BBC3 (PUMA) knockout HeLa cell lysate

Western blot - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)

Sanger Sequencing - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)

Sanger Sequencing - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)

Sanger Sequencing - Human BBC3 (PUMA) knockout HeLa cell lysate (AB256848)

주요 정보

세포 유형

Species or organism

조직

Knockout 검증

Mutation 정보

질병

보완 제품

Cell Lines & Lysates

Cell Lines & Lysates

Primary Antibodies

Assay Kits

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

추가 정보

Biological function summary

Pathways

품질 관리

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

제품 프로토콜

Product promise