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Human Pref-1/DLK1/FA1 Quantikine ELISA Kit

Catalog #: DPRF10
3 Citations Datasheet / COA / SDS

Discontinued Product

DPRF10 has been discontinued.
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Summary Precision Recovery Linearity Product Datasheets Preparation and Storage Background Related Research Areas

Human Pref-1/DLK1/FA1 Quantikine ELISA Kit Summary

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 (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
0.034 ng/mL
Assay Range
0.2 - 10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human Pref-1
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.

Product Summary

The Quantikine Human Pref-1 Immunoassay is a 4.5 hour sandwich ELISA designed to measure human Pref-1 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human Pref-1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Pref-1 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 Pref-1.

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, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 1.74 3.44 6.33 1.67 3.37 6.19
Standard Deviation 0.075 0.125 0.194 0.118 0.186 0.371
CV% 4.3 3.6 3.1 7.1 5.5 6

Recovery

The recovery of Pref-1 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 107 102-113
EDTA Plasma (n=4) 96 86-101
Heparin Plasma (n=4) 95 89-99
Serum (n=4) 99 91-107

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Pref-1 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: Pref-1/DLK1/FA1

Pref-1 (preadipocyte factor 1), also known as deltalike 1 (DLK1) and fetal antigen 1 (FA1), is a member of the Notch/Delta/Serrate protein family. It is a transmembrane protein that can be proteolyically cleaved to generate soluble forms. Pref-1 is believed to be a co-ligand for Notch signaling, enhancing or inhibiting Notch activation following binding of a ligand. It is expressed in numerous endocrine cells, and is highly expressed in preadipocytes, but not in mature adipocytes. In adipose tissue, Pref-1 inhibits adipogenesis and is often used as a preadipocyte marker.

Long Name:
Preadipocyte Factor-1/Protein delta Homolog 1/Fetal Antigen 1
Entrez Gene IDs:
8788 (Human); 13386 (Mouse)
Alternate Names:
delta-like 1 homolog (Drosophila); DLK; DLK1; DLK-1; FA1; FA1fetal antigen 1; pG2; pG2delta-like homolog (Drosophila); preadipocyte factor 1; Pref1; Pref-1; protein delta homolog 1; secredeltin; ZOG

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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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 for Human Pref-1/DLK1/FA1 Quantikine ELISA Kit

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.

3 Citations: Showing 1 - 3
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  1. Marrow fat and preadipocyte factor-1 levels decrease with recovery in women with anorexia nervosa.
    J. Bone Miner. Res., 2012-09-01;27(9):1864-71.
    Species: Human
    Sample Types: Serum
  2. DLK1, a serum marker for hepatoblastoma in young infants.
    Authors: Falix FA, Aronson DC, Lamers WH, Hiralall JK, Seppen J
    Pediatr Blood Cancer, 2011-12-16;59(0):743.
    Species: Human
    Sample Types: Serum
  3. Preadipocyte factor-1 is associated with marrow adiposity and bone mineral density in women with anorexia nervosa.
    Authors: Fazeli PK, Bredella MA, Misra M
    J. Clin. Endocrinol. Metab., 2009-10-22;95(1):407-13.
    Species: Human
    Sample Types: Serum

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