AB258146

Human PSMD10 (Gankyrin) knockout HeLa cell lysate

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PSMD10 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1.

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Lab

Western blot - Human PSMD10 (Gankyrin) knockout HeLa cell lysate (AB258146)

Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
Lane 2 : PSMD10 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate (20 ug)
Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (20 ug)

ab188315 was shown to specifically react with Gankyrin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265680 (knockout cell lysate ab258146) was used. Wild-type and Gankyrin knockout samples were subjected to SDS-PAGE. ab188315 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^oC at 1 in 1000 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.

All lanes:

Western blot - Anti-Gankyrin antibody [EPR14456(2)] (<a href='/ko/products/primary-antibodies/gankyrin-antibody-epr144562-ab188315'>ab188315</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

PSMD10 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PSMD10 (Gankyrin) knockout HeLa cell line (<a href='/ko/products/cell-lines/human-psmd10-gankyrin-knockout-hela-cell-line-ab265680'>ab265680</a>)

Lane 3:

K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate at 20 µg

Lane 4:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

Sanger Sequencing - Human PSMD10 (Gankyrin) knockout HeLa cell lysate (AB258146)

Sanger seq

Unknown

Sanger Sequencing - Human PSMD10 (Gankyrin) knockout HeLa cell lysate (AB258146)

Homozygous : 1 bp insertion in exon1

Lab

Western blot - Human PSMD10 (Gankyrin) knockout HeLa cell lysate (AB258146)

ab182576 was shown to specifically react with Gankyrin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265680 (knockout cell lysate ab258146) was used. Wild-type and Gankyrin knockout samples were subjected to SDS-PAGE. ab182576 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^oC at 1 in 1000 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.

All lanes:

Western blot - Anti-Gankyrin antibody [EPR14455] (<a href='/ko/products/primary-antibodies/gankyrin-antibody-epr14455-ab182576'>ab182576</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PSMD10 (Gankyrin) knockout HeLa cell lysate (ab258146) at 20 µg

Lane 3:

K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate at 20 µg

Lane 4:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

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, 1 bp insertion in exon1.

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":"ab258146-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB258146-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human PSMD10 knockout HeLa cell lysate", "number":"AB258146-CMP01", "productcode":"" } ] } } }

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특성 및 보관 정보

유전자명

PSMD10

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.

Gankyrin or PSMD10 is an oncogene with a molecular mass of around 25 kDa. It is a small heat-stable protein predominantly expressed in the cytoplasm of various tissues. The protein contains multiple ankyrin repeats that facilitate protein-protein interactions. It is overexpressed in many types of cancer tissues including liver breast and lung cancer. Gankyrin interacts with various cell cycle regulators and serves as an adaptor molecule enhancing its significance in cellular processes.

Biological function summary

Gankyrin enhances the degradation of the retinoblastoma protein (pRb) as part of the 26S proteasome complex. This function disrupts normal cell cycle control promoting cell proliferation. Gankyrin binds directly to the cyclin-dependent kinase 4 (CDK4) and MDM2 which regulate pRb and p53 tumor suppressor pathways. Overexpression of Gankyrin contributes to cellular transformation and oncogenesis by regulating cell cycle progression and apoptosis.

Pathways

Gankyrin participates actively in p53 and ubiquitin-proteasome pathways. The protein facilitates ubiquitination and degradation of key regulators including p53 and pRb altering the stability of these proteins. Gankyrin interacts with proteins like MDM2 and HDAC1 influencing their roles in tumor development through these pathways. Its presence in these pathways contributes to unchecked cell growth by evading normal tumor suppressor mechanisms.

Gankyrin has strong associations with cancer particularly in hepatocellular carcinoma and colorectal cancer. In these cancers it influences tumor progression and metastasis by altering the functions of proteins within the p53 pathway such as MDM2 and pRb. In hepatocellular carcinoma increased Gankyrin activity correlates with poor prognosis and aggressive tumor behavior while in colorectal cancer its interaction with these proteins may contribute to chemotherapy resistance.

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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

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 PSMD10 (Gankyrin) knockout HeLa cell lysate

Western blot - Human PSMD10 (Gankyrin) knockout HeLa cell lysate (AB258146)

Sanger Sequencing - Human PSMD10 (Gankyrin) knockout HeLa cell lysate (AB258146)

Western blot - Human PSMD10 (Gankyrin) knockout HeLa cell lysate (AB258146)

주요 정보

세포 유형

Species or organism

조직

Knockout 검증

Mutation 정보

질병

보완 제품

Primary Antibodies

Cell Lines & Lysates

Proteins & Peptides

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