Drug Names

Generic Name：Canine hepcidin(Hepc)ELISA Kit

Purpose

Our Canine hepcidin(Hepc) ELISA kit is to assay Hepc levels in Canine serum, plasma, culture media or any biological fluid.

Principle

This ELISA kit uses Sandwich-ELISA as the method. The Microelisa stripplate provided in this kit has been pre-coated with an antibody specific to Hepc. Standards or samples are added to the appropriate Microelisa stripplate wells and combined to the specific antibody. Then a Horseradish Peroxidase (HRP)- conjugated antibody specific for Hepc is added to each Microelisa stripplate well and incubated. Free components are washed away. The TMB substrate solution is added to each well. Only those wells that contain Hepc and HRP conjugated Hepc 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 Hepc. You can calculate the concentration of Hepc in the samples by comparing the OD of the samples to the standard curve.

 

Materials provided with the kit

	Materials provided with the kit	96 determinations	Storage
1	User manual	1	R.T.
2	Closure plate membrane	2	R.T.
3	Sealed bags	1	R.T.
4	Microelisa stripplate	1	2-8℃
5	Standard：10.8 ng/ml	0.5ml×1 bottle	2-8℃
6	Standard diluent	1.5ml×1 bottle	2-8℃
7	HRP-Conjugate reagent	6ml×1 bottle	2-8℃
8	Sample diluent	6ml×1 bottle	2-8℃
9	Chromogen Solution A	6ml×1 bottle	2-8℃
10	Chromogen Solution B	6ml×1 bottle	2-8℃
11	Stop Solution	6ml×1 bottle	2-8℃
12	wash  solution	20ml (30X)×1bottle	2-8℃

Sample preparation

1. 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.

2. 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.

3. Urine samples

Collect urine into aseptic tubes. 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. The preparation procedure of cerebrospinal fluid and pleuroperitoneal fluid is the same as that of urine sample.

4. 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

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