Human SMURF2 knockout HeLa cell line
- Advanced Validation
- What is this?
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SMURF2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 5 and 2 bp deletion in exon 5.
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
- WB
Lab
Western blot - Human SMURF2 knockout HeLa cell line (AB265231)
Lanes 1-3 : Merged signal (red and green). Green - ab53316 observed at 86 kDa. Red - loading control ab8245 observed at 36 kDa.
ab53316 Anti-SMURF 2 antibody [EP629Y3] 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 4°C 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 (ab265231)
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
false
- Sanger seq
Unknown
Sanger Sequencing - Human SMURF2 knockout HeLa cell line (AB265231)
Allele-1 : 2 bp deletion in exon 5.
- Sanger seq
Unknown
Sanger Sequencing - Human SMURF2 knockout HeLa cell line (AB265231)
Allele-2 : 1 bp insertion in exon 5.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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.
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
Handling procedures
Initial handling guidelines
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 guidelines
- 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.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
![Western blot - Anti-SMURF 2 antibody [EP629Y3] (AB53316)](https://content.abcam.com/products/images/smurf-2-antibody-ep629y3-ab53316--western-blot-img25486.png)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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