Table 1.
Validation of the Sensitive LH ELISA: Spike Recovery, Repeatability Precision, Direct Comparison with RIA, and Deconvolution and ApEN Analysis Parameters of Pulsatile LH Secretion from 10-Week-Old Male C57BL/6J Mice
| Whole Blood | Plasma | |
|---|---|---|
| Spike recovery, ng/mL mLH | ||
| 0.500 ng/mL | 87.87 ± 2.450 | 86.62 ± 1.220 |
| 0.125 ng/mL | 88.30 ± 2.580 | 88.53 ± 2.600 |
| 0.031 ng/mL | 84.93 ± 4.600 | 86.10 ± 7.290 |
| Repeatability precision, ng/mL mLH or % | ||
| Intra-assay CV | 0.250 ± 0.002 | 6.05 |
| Interassay CV | 0.220 ± 0.002 | 4.29 |
| Direct comparison with an established RIA | ||
| Slope (R2) | 1.192 ± 0.071 (0.921) | |
| Deconvolution and ApEn parameters, mean ± SEM (n = 8) | ||
| Total LH secretion, ng/mL/6 h | 13.9 ± 2.26 | |
| Pulsatile LH secretion rate, ng/mL/6 h | 5.86 ± 1.51 | |
| Mass of LH secreted/burst (MPP), ng/mL | 1.76 ± 0.14 | |
| Basal LH secretion rate, ng/mL/6 h | 2.64 ± 0.52 | |
| No. of pulses/6 h | 3.25 ± 0.59 | |
| Half-life, min | 10.4 ± 0.56 | |
| ApEn, 1, 0.35 | 0.46 ± 0.04 |
Abbreviation: CV, coefficient of variation. Recovery of LH in whole blood samples was quantified by spiking samples with 0.5, 0.125, and 0.031 ng/mL mLH. The intra- and inter-assay CVs were determined using the mLH standard. For pulsatile measures of LH secretion, samples were collected at 10-minute intervals between 7:00 am and 1:00 pm.