Human PDGFRA knockout SH-SY5Y cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
PDGFRA KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 67 bp deletion in exon 3. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
Alpha-type platelet-derived growth factor receptor, CD140 antigen-like family member A, CD140A, CD140a antigen, MGC74795, PDGF Receptor alpha, PDGF alpha chain, PDGF-R-alpha, PDGFR 2, PDGFRA, PDGFRA/BCR fusion, PGFRA_HUMAN, Platelet derived growth factor receptor, Platelet derived growth factor receptor 2, Platelet derived growth factor receptor alpha, Platelet derived growth factor receptor alpha polypeptide, RHEPDGFRA, Rearranged in hypereosinophilia platelet derived growth factor receptor alpha fusion protein
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human PDGFRA knockout SH-SY5Y cell line (AB275335)
ab203491 staining PDGFR alpha in wild-type SH-SY5Y cells (top panel) and PDGFRA knockout SH-SY5Y cells (bottom panel) (ab275335). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab203491 at 1μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
- WB
Lab
Western blot - Human PDGFRA knockout SH-SY5Y cell line (AB275335)
Lanes 1 - 2 : Merged signal (red and green). Green - ab203491 observed at 150 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab203491 was shown to react with PDGFR alpha in wild-type SH-SY5Y cells in Western blot with loss of signal observed in PDGFRA knockout cell line ab275335 (knockout cell lysate ab275522). Wild-type SH-SY5Y and PDGFRA knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab203491 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 ° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-PDGFR alpha antibody [EPR22059-270] (<a href='/en-us/products/primary-antibodies/pdgfr-alpha-antibody-epr22059-270-ab203491'>ab203491</a>) at 1/1000 dilution
Lane 1:
Wild-type SH-SY5Y cell lysate at 40 µg
Lane 2:
PDGFRA knockout SH-SY5Y cell lysate at 40 µg
Lane 2:
Western blot - Human PDGFRA knockout SH-SY5Y cell line (ab275335)
Predicted band size: 122 kDa
Observed band size: 150 kDa
false
- WB
Lab
Western blot - Human PDGFRA knockout SH-SY5Y cell line (AB275335)
Lanes 1 - 2 : Merged signal (red and green). Green - ab134123 observed at 180 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab134123 was shown to react with PDGFR alpha in wild-type SH-SY5Y cells in Western blot with loss of signal observed in PDGFRA knockout cell line ab275335 (knockout cell lysate ab275522). Wild-type SH-SY5Y and PDGFRA knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab134123 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 ° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-PDGFR alpha antibody [EPR5480] (<a href='/en-us/products/primary-antibodies/pdgfr-alpha-antibody-epr5480-ab134123'>ab134123</a>) at 1/1000 dilution
Lane 1:
Wild-type SH-SY5Y cell lysate at 40 µg
Lane 2:
PDGFRA knockout SH-SY5Y cell lysate at 40 µg
Lane 2:
Western blot - Human PDGFRA knockout SH-SY5Y cell line (ab275335)
Predicted band size: 122 kDa
Observed band size: 180 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human PDGFRA knockout SH-SY5Y cell line (AB275335)
Allele-1 : 67 bp deletion in exon 3
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 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.
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.
- These cells grow as a mixture of floating and adherent cells.
- Remove media containing floating cells and recover cells by centrifugation, detach cells using standard methods, combine with floating cells and transfer to a new culture flask.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be seeded at a density conducive to cell–cell communication to proliferate. If cells are seeded too sparsely, growth rate is reduced and cell death is high.
Culture medium
1:1 mixture of EMEM and F-12K + 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
PDGFR alpha mediates cell proliferation survival and differentiation especially in embryonic development and wound healing. The receptor often forms a complex by dimerizing with itself or with PDGFR beta to activate intracellular signaling cascades. This process is initiated upon ligand binding which triggers autophosphorylation on tyrosine residues. The receptor is also recognized in the detection of PDGFRA with assays such as PDGFRA ELISA.
Pathways
PDGFR alpha significantly affects the MAPK and PI3K/AKT signaling pathways which regulate cellular growth and survival signals. Through these pathways PDGFR alpha interacts with related proteins like SHP-2 and PI3K contributing to signal transduction involved in cellular responses. This role links PDGFR alpha to important cellular processes via its kinase activity and signal integration.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB203491
Anti-PDGFR alpha antibody [EPR22059-270]
BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha antibody [EPR22059-270] (AB203491)](https://content.abcam.com/products/images/pdgfr-alpha-antibody-epr22059-270-ab203491--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img418632.jpg)
- 4
- 5
![Immunocytochemistry/ Immunofluorescence - Anti-NG2 antibody [EPR23976-145] (AB275024)](https://content.abcam.com/products/images/ng2-antibody-epr23976-145-ab275024--immunocytochemistry-immunofluorescence-img184207.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