Natural and recombinant mouse and rat PDGF-BB. This assay cross-reacts 10.2% with recombinant Rat PDGF-AB, 5.3% with recombinant Human PDGF-AB, 64.7% with recombinant Human PDGF-BB, 38.1% with natural Human PDGF and 65.2% with natural Porcine PDGF.
Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity observed with 1 or more species tested.
Interference observed with 1 or more available related molecules.
The Quantikine Mouse/Rat PDGF-BB Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse/rat PDGF-BB in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant rat PDGF-BB and antibodies raised against recombinant human PDGF-BB. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse/rat PDGF-BB showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse/rat PDGF-BB.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates
Serum, EDTA Plasma, Heparin Plasma
The recovery of PDGF-BB spiked to three levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Mouse Cell Culture Supernates (n=8)
Mouse Heparin Plasma (n=5)
Rat Cell Culture Supernates (n=8)
Rat EDTA Plasma (n=5)
Rat Heparin Plasma (n=5)
Rat Serum (n=5)
To assess the linearity of the assay, samples spiked with or containing PDGF-BB in each matrix were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
The platelet-derived growth factor (PDGF) family of disulfide-linked dimeric proteins consists of four homodimeric proteins, PDGF-AA, PDGF-BB, PDGF-CC and PDGF-DD, and one heterodimeric protein, PDGF-AB. Members of this protein family act mainly on connective tissue. The dimeric isoforms of PDGF are expressed by different cell types and at different times during embryonic development. All PDGF isoforms are synthesized as inactive precursors and are processed to active forms that contain a PDGF/VEGF cysteine knot domain. Only PDGF-C and PDGF-D, also named spinal cord-derived growth factors SCDGF and SCDGF-B, also contain an N-terminal CUB domain. Mature PDGF dimers bind and induce the homo- or hetero-dimerization of two receptor tyrosine kinases (PDGF R alpha and R beta).
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
50 µL Assay Diluent
Add 50 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
100 µL Conjugate
Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
Aspirate and wash 5 times.
100 µL Substrate Solution
Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
100 µL Stop Solution
Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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