Human PDGF-DD Quantikine ELISA Kit DDD00: R&D Systems

Human PDGF-DD Quantikine ELISA Kit

(1 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (10 uL), Platelet-poor EDTA Plasma (50 uL), Platelet-poor Heparin Plasma (50 uL), Saliva (50 uL), Human Milk (50 uL)
  • Sensitivity
    3.51 pg/mL
  • Assay Range
    31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Saliva, Human Milk)
  • Specificity
    Natural and recombinant human PDGF-DD
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC140, Quantikine Immunoassay Control Set 725 for Human PDGF-DD - Please Inquire
Product Summary
The Quantikine Human PDGF-DD Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human PDGF-DD in cell culture supernates, serum, plasma, saliva, and human milk. It contains NS0-expressed recombinant human PDGF-DD and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human PDGF-DD 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 human PDGF-DD.

Precision
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, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Saliva, Human Milk
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean1765109861985591095
Standard Deviation3.6911.917.21638.555.3
CV%2.12.31.78.16.95.1

Recovery

The recovery of PDGF-DD spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 102 83-112
Platelet-poor EDTA Plasma (n=4) 93 83-109
Platelet-poor Heparin Plasma (n=4) 94 84-113
Serum (n=4) 101 86-112
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of PDGF-DD were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human PDGF-DD Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: PDGF-DD
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).
    • Long Name
      Platelet-derived Growth Factor DD
    • Entrez Gene IDs
      80310 (Human);
    • Alternate Names
      PDGFDD; PDGF-DD;
    Related Research Areas
    Assay Procedure
    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.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    1 Citations: Showing 1 - 1

    1. PDGF-D expression is down-regulated by TGFbeta in fibroblasts.
      Authors: Charni Chaabane S, Coomans de Brachene A, Essaghir A, Velghe A, Lo Re S, Stockis J, Lucas S, Khachigian L, Huaux F, Demoulin J
      PLoS ONE, 2014;9(10):e108656.
      Species: Human
      Sample Type: Cell Culture Supernates
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 725 for Human PDGF-DD

    Ctrl QC140
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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