AB258146

Human PSMD10 (Gankyrin) knockout HeLa cell lysate

Name: Human PSMD10 (Gankyrin) knockout HeLa cell lysate (ab258146) | Abcam
Brand: Abcam Limited
SKU: AB258146

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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.

3 Images

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='/en-us/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='/en-us/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='/en-us/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

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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='/en-us/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:

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='/en-us/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:

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

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1.

Disease

Adenocarcinoma

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Reactivity data

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

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Product details

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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What's included?

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Properties and storage information

Gene name

PSMD10

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Shipped at conditions

Ambient - Can Ship with Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

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Supplementary information

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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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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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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)

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

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

Key facts

Cell type

Species or organism

Tissue

Knockout validation

Mutation description

Disease

Complementary Products

Cell Lines & Lysates

Proteins & Peptides

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Product promise