PFN2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2
Alternative names
D3S1319E, PFL, PFN 2, PROF2_HUMAN, Profilin II, Profilin-2
Recommended products
PFN2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- PFN2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- Zygosity
- Homozygous
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, 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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Profilin 2 also known as PFN2 is an actin-binding protein with an important role in the regulation of the actin cytoskeleton. With a molecular weight of around 14 kDa it is expressed in tissues such as the brain and muscle where it contributes to dynamic cellular processes such as cell movement and shape maintenance. PFN2 binds not only to actin but also to phosphatidylinositol 45-bisphosphate mediating the sequestration and availability of actin monomers essential for actin polymerization.
- Biological function summary
Profilin 2 regulates actin dynamics essential for cellular processes like membrane trafficking and signal transduction. It is not typically known to be part of an extensive protein complex but interacts with other proteins including members of the Arp2/3 complex to promote actin filament nucleation and branching. This regulation impacts processes like vesicle transport particularly in neuronal cells where Profilin 2 influences synaptic function and plasticity.
- Pathways
Actin dynamics controlled by Profilin 2 play significant roles in both the Rho GTPase and PI3K/AKT signaling pathways. In these pathways Profilin 2 interfaces with Rho-related proteins to modulate the actin cytoskeleton impacting cell adhesion and movement. Its interaction with PI3K elements contributes to cell growth and survival signals aligning Profilin 2's functions with cellular responses to environmental cues.
- Associated diseases and disorders
Abnormalities in Profilin 2 expression or function have been linked to neurological disorders such as schizophrenia and neurodegenerative diseases like Amyotrophic Lateral Sclerosis (ALS). In these conditions Profilin 2’s interactions with cytoskeletal proteins such as cofilin and tau could disrupt normal neuronal functions and synaptic stability. Its deregulation resulting in misbalanced actin dynamics contributes to the pathology observed in affected neuronal circuits.
Product promise
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2 product images
Western blot - Human PFN2 (Profilin 2) knockout HeLa cell line (ab265833)
False colour image of Western blot: Anti-Profilin 2 antibody [EPR11030] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-Profilin 2 antibody [EPR11030] ab174322 was shown to bind specifically to Profilin 2. A band was observed at 13 kDa in wild-type HeLa cell lysates with no signal observed at this size in PFN2 knockout cell line ab265833 (knockout cell lysate Human PFN2 (Profilin 2) knockout HeLa cell lysate ab258108). To generate this image wild-type and PFN2 knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-Profilin 2 antibody [EPR11030] (Anti-Profilin 2 antibody [EPR11030] ab174322) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: PFN2 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human PFN2 (Profilin 2) knockout HeLa cell line (ab265833)
Lane 3: SH-SY5Y cell lysate at 20 µg
Lane 4: Daudi cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 13 kDa
Sanger Sequencing - Human PFN2 (Profilin 2) knockout HeLa cell line (ab265833)
Homozygous: 1 bp insertion in exon 2.
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