Various Species Prostaglandin F ELISA Kit, Lot 21AO-637 [Cancer Immune Checkpoint Assay Kit] (CAT#: IOK-05-P548)

18.75 pg/mL

50 μL

Pre-coated

1.Bring all reagents to room temperature (18-25 °C) before use.
2.Wash Buffer: Dilute 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water to prepare 750 mL of Wash Buffer.
3.Standard working solution: Add 1.0 mL of Reference Standard &Sample Diluent, let it stand for 10 min and invert it gently several times. After it dissolves fully, mix it thoroughly with a pipette. This reconstitution produces a working solution of 200pg/mL.
4.Biotinylated Detection Ab working solution: Centrifuge the Concentrated Biotinylated Detection Ab at 800xg for 1 min, then dilute the 100x Concentrated Biotinylated Detection Ab to 1x working solution with Biotinylated Detection Ab Diluent.
5.HRP Conjugate working solution: Calculate the required amount before the experiment.

1.Add the Standard working solution to the first two columns: Each concentration of the solution is added in duplicate, to one well each, side by side (100 μL for each well).Cover the plate with the sealer provided in the kit. Incubate for 90 min at 37 °C.
2.Remove the liquid out of each well, do not wash. Immediately add 100 μL of Biotinylated Detection Ab working solution to each well. Cover with the Plate sealer. Gently mix up. Incubate for 1 hour at 37 °C.
3.Aspirate or decant the solution from each well, add 350 μL of wash buffer to each well. Soak for 1~2 min and aspirate or decant the solution from each well and pat it dry against clean absorbent paper. Repeat this wash step 3 times.
4.Add 100 μL of HRP Conjugate working solution to each well. Incubate for 30 min at 37 °C.
5.Aspirate or decant the solution from each well, repeat the wash process for five times as conducted in step 3.
6.Add 90 μL of Substrate Reagent to each well. Cover with a new plate sealer. Incubate for about 15 min at 37 °C. Protect the plate from light.
7.Add 50 μL of Stop Solution to each well.
8.Determine the optical density (OD value) of each well at once with a micro-plate reader set to 450 nm.

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level PGF were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level PGF were tested on 3 different plates, 20 replicates in each plate.
Both intra-CV and inter-CV are < 10 %.

4 °C,-20 °C

An unopened kit can be stored at 2-8°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

6 months

An unopened kit can be stored at 2-8°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

For Research Use only

Prostaglandin F (PG-F)

1.Add 50 μL standard or sample to each well. Immediately add 50 μL Biotinylated Detection Ab to each well. Incubate for 45 min at 37 °C.
2.Aspirate and wash 3 times.
3.Add 100 μL HRP Conjugate to each well. Incubate for 30 min at 37 °C.
4.Aspirate and wash 5 times.
5.Add 90μL Substrate Reagent. Incubate 15 min at 37 °C.
6.Add 50 μL Stop Solution. Read at 450nm immediately.
7.Calculation of results.