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PDGFRA KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 67 bp deletion in exon 3.

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Images

Western blot - Human PDGFRA knockout SH-SY5Y cell line (AB275335), expandable thumbnail
  • Western blot - Human PDGFRA knockout SH-SY5Y cell line (AB275335), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Human PDGFRA knockout SH-SY5Y cell line (AB275335), expandable thumbnail
  • Sanger Sequencing - Human PDGFRA knockout SH-SY5Y cell line (AB275335), expandable thumbnail

Key facts

Cell type
SH-SY5Y
Species or organism
Human
Tissue
Bone marrow
Form
Liquid
Knockout validation
Immunocytochemistry, Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 67 bp deletion in exon 3

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

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PDGFRA KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 67 bp deletion in exon 3.

Key facts

Cell type
SH-SY5Y
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 67 bp deletion in exon 3
Disease
Neuroblastoma
Concentration
Loading...

Properties

Gene name
PDGFRA
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Immunocytochemistry, Sanger Sequencing, Western blot
Zygosity
Homozygous

Quality control

STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level
EU: 1 US: 1
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.
  • 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.

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

Recommended control: Human wild-type SHSY-5Y cell line (ab275475). 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 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

PDGFR alpha also known as platelet-derived growth factor receptor alpha is a cell surface receptor tyrosine kinase with a mass of approximately 123 kDa. It plays a mechanical role in transmitting signals across the cell membrane initiating cellular responses. PDGFR alpha is widely expressed in mesenchymal cells including fibroblasts and smooth muscle cells. Researchers frequently employ techniques such as PDGFRA IHC and PDGFRA staining to study the localization and expression levels in tissues.

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.

Associated diseases and disorders

PDGFR alpha is implicated in conditions such as gastrointestinal stromal tumors (GISTs) and idiopathic pulmonary fibrosis (IPF). It connects to proteins such as KIT in GISTs where mutations cause aberrant receptor signaling leading to tumorigenesis. Aberrant PDGFR alpha signaling also associates with IPF a disorder characterized by increased fibroblast proliferation and fibrotic tissue formation. These connections make PDGFR alpha a significant target for research and therapeutic intervention.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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