GSN 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 deletion in exon 6 and 20 bp deletion in exon 6.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 6 and 20 bp deletion in exon 6
Alternative names
ADF, AGEL, Actin-depolymerizing factor, Brevin, DKFZp313L0718, GELS_HUMAN, Gelsolin, Gsn
Recommended products
GSN 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 deletion in exon 6 and 20 bp deletion in exon 6.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 6 and 20 bp deletion in exon 6
- Antibiotic resistance
- Puromycin 1µg/mL
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- GSN
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Gelsolin is a multifunctional actin-binding protein that exists in two main isoforms: cytoplasmic and plasma. It has a molecular mass of approximately 82 kDa. Gelsolin is expressed in a wide range of tissues including muscle epithelial and nerve cells. The protein's main mechanical function involves severing actin filaments which helps to regulate actin polymerization and depolymerization. Researchers sometimes refer to gelsolin as 'GSN'. The presence of the protein helps maintain the dynamic organization of the cytoskeleton by interacting with actin filaments.
- Biological function summary
This protein acts in cell movement apoptosis and signal transduction. Gelsolin contributes to the remodeling of the actin cytoskeleton by promoting the disassembly of actin filaments allowing cells to change shape and move. Although it operates independently gelsolin’s activity can be influenced by calcium ions and phosphoinositides leading to its participation in various cellular activities. Gelsolin plays a role in platelet activation and may regulate cellular responses through association with the cytoskeletal complex.
- Pathways
Gelsolin participates in the regulation of the actin cytoskeleton and apoptotic pathways. Within these pathways gelsolin interacts with proteins like cofilin to modulate actin filament dynamics. In apoptotic pathways it collaborates by altering the cytoskeletal architecture impacting cell death mechanisms. Gelsolin modifications in these pathways have implications for cellular processes like cell migration and apoptosis bringing it into relation with proteins such as caspases.
- Associated diseases and disorders
Gelsolin is linked to conditions such as gelsolin amyloidosis and cancer metastasis. Gelsolin amyloidosis results from mutations in the gelsolin gene which lead to the formation of amyloid fibrils causing dysfunction in nerves eyes and skin. In cancer altered gelsolin expression can influence tumor progression and metastasis through its role in cell motility. Related proteins such as actin and caspases could also play a part in these conditions either through direct interaction or through pathways in which gelsolin functions.
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6 product images
Western blot - Human GSN (Gelsolin) knockout HeLa cell line (ab265201)
Anti-Gelsolin antibody [EPR1942] ab109014 was shown to react with GSN in wild-type HeLa cells in Western blot with loss of signal observed in GSN knockout cell line ab265201 (GSN knockout cell lysate Human GSN (Gelsolin) knockout HeLa cell lysate ab257204). Wild-type HeLa and GSN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-Gelsolin antibody [EPR1942] ab109014 overnight at 4 ° at a 1/20000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 1/5000 before imaging. These data were provided by YCharOS Inc. an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-Gelsolin antibody [EPR1942] (Anti-Gelsolin antibody [EPR1942] ab109014) at 1/20000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: GSN knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human GSN (Gelsolin) knockout HeLa cell line (ab265201)
Performed under reducing conditions.
Predicted band size: 86 kDa
Western blot - Human GSN (Gelsolin) knockout HeLa cell line (ab265201)
Anti-Gelsolin antibody ab74420 was shown to react with GSN in wild-type HeLa cells in Western blot with loss of signal observed in GSN knockout cell line ab265201 (GSN knockout cell lysate Human GSN (Gelsolin) knockout HeLa cell lysate ab257204). Wild-type HeLa and GSN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-Gelsolin antibody ab74420 overnight at 4 ° at a 1/20000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 1/5000 before imaging. This data was kindly provided by the YCharOS Inc. an open science company with the mission of characterizing every commercially available antibody reagent. abcam are working with YCharOS to support their mission of antibody characterisation using knockout cell lines.
All lanes: Western blot - Anti-Gelsolin antibody (Anti-Gelsolin antibody ab74420) at 1/20000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: GSN knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human GSN (Gelsolin) knockout HeLa cell line (ab265201)
Performed under reducing conditions.
Predicted band size: 86 kDa
Western blot - Human GSN (Gelsolin) knockout HeLa cell line (ab265201)
Anti-Gelsolin antibody [GS-2C4] ab11081 was shown to react with GSN in wild-type HeLa cells in Western blot with loss of signal observed in GSN knockout cell line ab265201 (GSN knockout cell lysate Human GSN (Gelsolin) knockout HeLa cell lysate ab257204). Wild-type HeLa and GSN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-Gelsolin antibody [GS-2C4] ab11081 overnight at 4 ° at a 1/200 dilution. Blots were incubated with goat anti-mouse HRP secondary antibodies at 1/5000 before imaging. This data was kindly provided by the YCharOS Inc. an open science company with the mission of characterizing every commercially available antibody reagent. abcam are working with YCharOS to support their mission of antibody characterisation using knockout cell lines.
All lanes: Western blot - Anti-Gelsolin antibody [GS-2C4] (Anti-Gelsolin antibody [GS-2C4] ab11081) at 1/200 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: GSN knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human GSN (Gelsolin) knockout HeLa cell line (ab265201)
Performed under reducing conditions.
Predicted band size: 86 kDa
Cell Culture - Human GSN (Gelsolin) knockout HeLa cell line (ab265201)
Representative images of GSN knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Sanger Sequencing - Human GSN (Gelsolin) knockout HeLa cell line (ab265201)
Allele-2: 1 bp deletion in exon 6.
Sanger Sequencing - Human GSN (Gelsolin) knockout HeLa cell line (ab265201)
Allele-1: 20 bp deletion in exon 6.
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