Human PFN2 (Profilin 2) knockout HeLa cell line
- Advanced Validation
- What is this?
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PFN2 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
D3S1319E, PFL, PFN 2, PROF2_HUMAN, Profilin II, Profilin-2
- WB
Lab
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] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-Profilin 2 antibody [EPR11030] (<a href='/en-us/products/primary-antibodies/profilin-2-antibody-epr11030-ab174322'>ab174322</a>) 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
Predicted band size: 15 kDa
Observed band size: 13 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PFN2 (Profilin 2) knockout HeLa cell line (AB265833)
Homozygous : 1 bp insertion in exon 2.
Reactivity data
Product details
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
Zygosity
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Immunoprecipitation - Anti-Profilin 2 antibody [EPR11030] (AB174322)](https://content.abcam.com/products/images/profilin-2-antibody-epr11030-ab174322--immunoprecipitation-img74005.jpg)
Product promise
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