1. The total assay time is about 20 hours.
2. The kit measures shedded HLA-G1 and HLA-G5 in EDTA plasma, amniotic fluid, or cell culture supernatant.
3. Calibrator is human native protein.
4. Assay format is 96 wells.
5. Components of the kit are provided ready to use, concentrated or lyophilized.Typical analytical data of sHLA-G ELISA are presented in this chapter Sensitivity
Limit of Detection (LOD), defined as concentration of analyte giving absorbance higher than mean absorbance of blank*plus three standard deviations of the absorbance of blank: Ablank+ 3xSDblank, is calculated from the real sHLA-G values in wells and is 0.6 Units/ml.
*Dilution Buffer is pipetted into blank wells. Limit of assay
Samples with absorbances exceeding the absorbance of the highest standard should be measured again with higher dilution. The final concentration of samples calculated from the standard curve must be multiplied by the respectivedilution factor.Presented results are multiplied by respective dilution factor Precision
Intra-assay (Within-Run) (n=8) for EDTA plasma samples diluted using the Dilution Buffer 2:
Inter-assay (Run-to-Run) (n=5) for EDTA plasma samples diluted using the Dilution Buffer 2:Spiking Recovery
EDTA plasma samples were spiked with different amounts of sHLA-G and assayed (Dilution Buffer 2 was used as a diluent).Linearity
EDTA plasma samples were serially diluted with Dilution Buffer 2 and assayed. Stability of samples stored at 2-8°C
A significant decline in concentration of sHLA-G was observed in EDTA plasma after 7 days when stored at 2-8°C. Therefore, we strongly recommend to store the samples at -20°C, or preferably at -70°C for long-term storage. Effect of Freezing/Thawing
Significant changes in concentration of sHLA-G wereobserved in EDTA plasma samples after repeated freeze/thaw cycles. Therefore, it is strongly recommended to avoid unnecessary repeated freezing/thawing of the samples. Reference range
It is recommended that each laboratory include its own panel of control samples in the assay. Each laboratory should establish its own normal andpathological reference ranges for sHLA-G levels with the assay.