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No .

E-EL-0033c

ST/5-HT(Serotonin/5-Hydroxytryptamine) ELISA Kit

Item No:

E-EL-0033c

Chinese Name:

Alias:

5HT, Enteramine, Thrombocytin, Thrombotonin, 3-(β-Aminoethyl)-5-hydroxyindole

Keywords:

ELISA kits

Classification:

Uncoated ELISA kit

Retail Price

¥ 1650

Market Price

¥ 1650

The kit is used for the in vitro quantitative determination of ST/5-HT concentrations in serum, plasma or other relevant biological fluids.

Spec
- 48T
- 96T
- 96T*5

Quantity

Inventory surplus

2997

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Describe

Use

The kit is used for the in vitro quantitative determination of ST/5-HT concentrations in serum, plasma or other relevant biological fluids.

Detection Principle

This kit utilizes a competitive ELISA method. ST/5-HT antigen is encapsulated with ST/5-HT on an enzyme labeled plate. During the experiment, ST/5-HT in the sample or standard competes with the encapsulated ST/5-HT for the binding site on the biotin labeled anti-ST/5-HT monoclonal antibody, and the free component is washed away. Horseradish peroxidase-labeled affinity hormone is added, biotin binds specifically to affinity hormone to form an immune complex, and the free component is washed away. Add the chromogenic substrate (TMB), TMB shows blue color catalyzed by horseradish peroxidase, and turns into yellow color after adding the termination solution. The OD value was measured at 450 nm with an enzyme marker, and there was an inverse ratio between the concentration of ST/5-HT and the OD450 value, and the concentration of ST/5-HT in the sample was calculated by plotting the standard curve.

Reaction type	competition law
Specification	96T
Reaction time	2.0h
Reactivity	common (use)
Detection method	Colormetric
Detection range	15.63-1000ng/mL
Sensitivity	9.38 ng/mL
Sample Volume	50μL
Sample Type	Serum, plasma or other biological fluids

Idiosyncrasy

Detects ST/5-HT in samples with no significant cross-reactivity with other related proteins.

Repeatable

The coefficients of variation within plates and between plates were all <10%.

Typical data

The OD values of the standard curve will vary due to different experimental operating conditions (e.g., operator, pipetting technique, plate washing technique and stabilization conditions, etc.). The following data and curves are for reference only, and the experimenter needs to establish the standard curve according to his/her own experiment.

(ng/mL)	O.D	Average
1000	0.402 0.426	0.414
500	0.528 0.532	0.53
250	0.73 0.722	0.726
125	1.004 1.018	1.011
62.5	1.355 1.343	1.349
31.25	1.674 1.654	1.664
15.63	1.899 1.901	1.9
0	2.217 2.221	2.219

Precision

Intra-plate precision: Low, medium and high concentration samples were tested 20 times on 1 plate.
　　Inter-plate precision: low concentration, medium concentration and high concentration samples were tested 20 times on 3 plates.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	51.20	141.30	362.70	54.70	132.00	351.00
Standard deviation	3.50	5.90	15.60	3.10	6.50	16.10
C V (%)	6.84	4.18	4.30	5.67	4.92	4.59

Recovery rate

A known concentration of the target protein was added to five different samples and recovery experiments were done to obtain a range of recoveries and an average recovery.

Sample Type	Range (%)	Average Recovery (%)
Serum (n=8)	95-107	100
EDTA plasma (n=8)	86-99	92
Cell culture media (n=8)	91-105	98

Linearity

A recovery experiment was performed by adding known concentrations of target protein to 5 samples to obtain a range of recoveries and average recoveries. Dilute the 5 samples 2x, 4x, 8x and 16x to obtain the recovery range and average recovery.

		Serum (n=5)	EDTA plasma (n=5)	Cell culture media (n=5)
1:2	Range (%)	90-104	86-99	97-112
1:2	Average (%)	97	92	104
1:4	Range (%)	83-96	92-103	96-108
1:4	Average (%)	90	98	102
1:8	Range (%)	84-97	86-99	93-108
1:8	Average (%)	90	92	101
1:16	Range (%)	84-97	94-107	94-109
1:16	Average (%)	90	99	100

Kit Composition and Storage

Unopened kits can be stored at 2-8°C for one week; if the kit is to be used after one week, unpack the kit and store the components separately according to the conditions in the table below.

Chinese name	English name	Specification	Storage conditions
ELISA酶标板(可拆卸)	Micro ELISA Plate(Dismountable)	8 holes×12 strips	-20℃, can be stored for 6 months
冻干标准品	Reference Standard	2 sticks
浓缩生物素化抗体(100×)	Concentrated Biotinylated Detection Ab	1 x 120 μL
浓缩HRP酶结合物(100×)	Concentrated HRP Conjugate	1 x 120 μL	-20℃ (avoid light), can be stored for 6 months
标准品&样品稀释液	Reference Standard & Sample Diluent	1 bottle 20 mL	2-8℃. Can be stored for 6 months
生物素化抗体稀释液	Biotinylated Detection Ab Diluent	1 bottle 14 mL
酶结合物稀释液	HRP Conjugate Diluent	1 bottle 14 mL
浓缩洗涤液(25×)	Concentrated Wash Buffer (25×)	1 bottle 30 mL
底物溶液(TMB)	Substrate Reagent	1 bottle 10 mL	2-8℃(avoid light)
反应终止液	Stop Solution	1 bottle 10 mL	2-8℃
封板覆膜	Plate Sealer	5 sheets
产品说明书	Product Description	1 sheet
质检报告	Certificate of Analysis	1 sheet

Note: All reagent bottle caps must be screwed on tightly to prevent evaporation and microbial contamination.
Reagent volumes are based on the actual shipping version of the instructions. Reagents may be dispensed in a slightly larger volume than indicated on the label, so please measure rather than pour when using.

Self-contained items required for testing

•1. Enzyme Labeling Instrument (450nm wavelength filter)

•2. High-precision pipettes, EP tubes and disposable tips: 0.5-10 μL, 2-20 μL, 20-200 μL, 200-1000 μL

•3.37℃ thermostat, double-distilled water or deionized water

•4. Pipette

Procedure

	1. Add 100μL of standard working solution or sample into the corresponding plate wells and incubate at 37℃ for 90 minutes.
	2. Discard the liquid in the plate and wash the plate 3 times.
	3. Add 100μL of HRP enzyme conjugate working solution to each well, incubate at 37℃ for 30 minutes, discard the liquid in the plate, and wash the plate 5 times.
	4. Add 90μL of substrate solution to each well and incubate at 37℃ for 15 minutes.
	5. Add 50μL of termination solution to each well.
	6. Immediately read at 450nm and process the data.