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AB257691

Human SMURF2 knockout HeLa cell lysate

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SMURF2 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 exon5 and 2 bp deletion in exon5.

View Alternative Names

E3 ubiquitin-protein ligase SMURF2, EC 6.3.2.-, MGC138150, SMAD ubiquitination regulatory factor 2, SMAD-specific E3 ubiquitin-protein ligase 2, SMUF2_HUMAN, Smad specific E3 ubiquitin ligase 2, Smurf2, Ubiquitin protein ligase SMURF2, hSMURF2

3 Images
Western blot - Human SMURF2 knockout HeLa cell lysate (AB257691)
  • WB

Lab

Western blot - Human SMURF2 knockout HeLa cell lysate (AB257691)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : SMURF2 knockout HeLa cell lysate (20 ug)
Lane 3 : SH-SY5Y cell lysate (20 ug)

ab53316 was shown to specifically react with SMURF 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265231 (knockout cell lysate ab257691) was used. Wild-type and SMURF 2 knockout samples were subjected to SDS-PAGE. ab53316 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC 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-SMURF 2 antibody [EP629Y3] (<a href='/en-us/products/primary-antibodies/smurf-2-antibody-ep629y3-ab53316'>ab53316</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SMURF2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SMURF2 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-smurf2-knockout-hela-cell-line-ab265231'>ab265231</a>)

Lane 3:

SH-SY5Y 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: 86 kDa

Observed band size: 86 kDa

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Sanger Sequencing - Human SMURF2 knockout HeLa cell lysate (AB257691)
  • Sanger seq

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Sanger Sequencing - Human SMURF2 knockout HeLa cell lysate (AB257691)

Allele-1 : 2 bp deletion in exon5

Sanger Sequencing - Human SMURF2 knockout HeLa cell lysate (AB257691)
  • Sanger seq

Unknown

Sanger Sequencing - Human SMURF2 knockout HeLa cell lysate (AB257691)

Allele-2 : 1 bp insertion in exon5

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 exon5 and 2 bp deletion in exon5.

Disease

Adenocarcinoma

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Reactivity data

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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
SMURF2
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.

SMURF2 also known as SMAD specific E3 ubiquitin protein ligase 2 functions as an E3 ubiquitin ligase involved in protein ubiquitination. It weighs approximately 85 kDa and is part of the NEDD4 family of E3 ligases. SMURF2 facilitates the transfer of ubiquitin from an E2 conjugating enzyme to specific substrate proteins marking them for degradation via the proteasome pathway. This protein is widely expressed in various tissues including brain kidney and liver indicating its essential role in different cellular processes.
Biological function summary

SMURF2 regulates many cell signaling pathways including the TGF-beta/Smad pathway. It associates with SMAD proteins to control their stability and activity acting as a negative regulator to modulate TGF-beta signaling. By promoting the ubiquitination and degradation of receptor-regulated SMADs (R-SMADs) SMURF2 impacts cellular responses to growth factors. This regulatory mechanism is important for maintaining cellular proliferation differentiation and apoptosis.

Pathways

The function of SMURF2 significantly influences the TGF-beta and BMP signaling pathways both critical in controlling cell growth and differentiation. These pathways involve interactions with other proteins like SMAD7 and SMAD2. SMURF2 interacts with SMAD7 an inhibitory SMAD to regulate Smad-dependent transcriptional responses ensuring the fine-tuning of signal transduction. These pathways' balance and regulation are vital for appropriate cellular function and response.

SMURF2 has connections to cancer and fibrosis. Aberrant regulation of SMURF2 contributes to tumorigenesis where its altered expression levels can lead to unrestrained cell growth and cancer progression particularly in breast and prostate cancers. Additionally SMURF2 is involved in fibrosis with its dysregulation affecting extracellular matrix deposition. In both cancer and fibrosis SMURF2 interacts with TGF-beta and other related proteins highlighting its involvement in disease progression and sustaining pathological states.
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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

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Primary Antibodies

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Product promise

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