Human Leptin Quantikine QuicKit ELISA

Catalog #: QK398 Datasheet

Catalog #	Availability	Size / Price	Qty
QK398

Control Products Available

QC264: Human Leptin Quantikine QuicKit Control Group 264

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All Leptin/OB ELISAs

Product Details

Procedure

FAQs

Supplemental Products

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Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human Leptin Quantikine QuicKit ELISA Summary

Assay Length

80 minutes

Sample Type & Volume Required Per Well

Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)

Sensitivity

1.03 pg/mL

Assay Range

15.6 - 1,000 pg/mL (Serum, EDTA Plasma, Heparin Plasma)

Specificity

Natural and recombinant human Leptin.

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^® QuicKit™ Human Leptin Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human Leptin levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human Leptin and antibodies raised against the recombinant protein. Results obtained using natural human Leptin showed linear curves that were parallel to the standard curves obtained using the QuicKit™ standards. These results indicate that this kit can be used to determine relative mass values for natural human Leptin.

Precision

Intra-Assay Precision (Precision within an assay) Two samples of known concentration were tested twenty times on one plate to assess intra-assay precision

Inter-Assay Precision (Precision between assays) Two samples of known concentration were tested in ten separate assays to assess inter-assay precision. Assays were performed by at least three technicians

Serum, EDTA Plasma, Heparin Plasma

	Intra-Assay Precision		Inter-Assay Precision
Sample	1	2	1	2
n	20	20	10	10
Mean (pg/mL)	134	709	130	709
Standard Deviation	2.84	15.8	11.9	19.7
CV%	2.1	2.2	9.2	2.8

Recovery

The recovery of human Leptin spiked to three levels in samples throughout the range of the assay was evaluated.

Sample Type	Average % Recovery	Range %
Cell Culture Media (n=4)	89	80-106

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Leptin were diluted with calibrator diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.

Scientific Data

N/A

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Human Leptin ELISA Standard Curve

ELISA

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Human Leptin QuicKit Spiked Recovery Competitor Comparison Leptin is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Serum recovery is 74% compared to 70% for the top competitor. EDTA plasma recovery is 84% compared to 69% for the top competitor. Heparin plasma recovery is 88% compared to 77% for the top competitor. In spike and recovery experiments, natural samples are spiked with the recombinant target analyte of interest to identify interference caused by sample matrices.

ELISA

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Human Leptin QuicKit Linearity Competitor Comparison Samples containing and/or spiked with high concentrations of Leptin in various matrices and diluted with appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay. The linearity is between 99%-106% compared to 71%-122% for the top competitor.

Product Datasheets

Product Datasheet

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 2 - 8 °C. Do not use past expiration date.

Background: Leptin/OB

Leptin is a product of the mouse obese gene. Mice with mutations in the obese gene that block the synthesis of leptin are obese and diabetic, and have reduced activity, metabolism, and body temperature. Rat leptin shares approximately 96% and 82% sequence identity with the mouse and human protein, respectively. The expression of leptin mRNA is restricted to adipose tissue.

Entrez Gene IDs:

3952 (Human); 16846 (Mouse); 25608 (Rat)

Alternate Names:

FLJ94114; LEP; leptin (murine obesity homolog); leptin (obesity homolog, mouse); Leptin; OB; Obese protein; obese, mouse, homolog of; Obesity factor; OBOBS

Related Research Areas

Assay Procedure

These assays utilize an anti-tag coated microplate. Standards, samples, and controls are added to plate, followed by subsequent addition of an antibody cocktail. Following a 1 hour incubation, plates are washed and substrate is added and incubated for 20 minutes. Stop solution is then added and plates are read using a standard microplate reader.

QuicKit ELISA assay procedure