Drug Generic Name: Human heme oxygenase 1,HO-1 ELISA Kit

Purpose

Our Human heme oxygenase 1,HO-1 ELISA Kit is to assay HO-1 levels in Human serum, plasma, culture media or any biological fluid.

Principle

This ELISA kit uses Sandwich-ELISA as the method. The Micro Elisa strip plate provided in this kit has been pre-coated with an antibody specific to HO-1. Standards or samples are added to the appropriate Micro Elisa strip plate wells and combined to the specific antibody. Then a Horseradish Peroxidase (HRP)-conjugated antibody specific for HO-1 is added to each
Micro Elisa strip plate well and incubated. Free components are washed away. The TMB substrate solution is added to each well. Only those wells that contain HO-1 and HRP conjugated HO-1 antibody will appear blue in color and then turn yellow after the addition of the stop solution. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm. The OD value is proportional to the concentration of HO-1. You can calculate the concentration of HO-1 in the samples by comparing the OD of the samples to the standard curve.

Sample Preparation

Serum Preparation: After collection of the whole blood, allow the blood to clot by leaving it undisturbed at room temperature. This usually takes 10-20 minutes. Remove the clot by centrifuging at 2,000-3,000 rpm for 20 minutes. If precipitates appear during reservation, the sample should be centrifugated again.

Plasma Preparation: Collect the whole blood into tubes with anticoagulant (EDTA or citrate). After incubated at room temperature for 10-20 minutes, tubes are centrifugated for 20 min at 2,000-3,000 rpm. Collect the supernatant carefully as plasma samples. If precipitates appear during reservation, the sample should be centrifugated again.

Urine Samples: Collect urine into aseptic tubes. Collect the supernatant carefully after centrifuging for 20min at 2,000-3,000 rpm. If precipitates appear during reservation, the sample should be centrifugated again. The preparation procedure of cerebrospinal fluid and pleuroperitoneal fluid is the same as that of urine sample.

Cell Samples: If you want to detect the secretions of cells, collect culture supernatant into aseptic tubes. Collect the supernatant carefully after centrifuging for 20 min at 2,000-3,000 rpm. If you want to detect intracellular components, dilute the cells to 1X100/ml with PBS (pH 7.2-7.4). The cells were destroyed to release intracellular components by repeated freezing and thawing. Collect the supernatant carefully after centrifuging for 20 min at 2,000-3,000 rpm. If precipitates appear during reservation, the sample should be centrifugated again.

Tissue Samples: Tissue samples are cut, weighed, frozen in liquid nitrogen and stored at -80℃ for future use. The tissue samples were homogenized after adding PBS (pH 7.4). Samples should be operated at 4℃ . Collect the supernatant carefully after centrifuging for 20 min at 2,000-3,000 rpm. Aliquot the supernatant for ELISA assay and future use.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Human HO-1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human HO-1 were tested on 3 different plates, 8 replicates in each plate.

CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%