Serotonin ELISA kit

Sample

Urine and serum

Species Reactivity

Human

Detection Method

ELISA

Intended Use

Enzyme Immunoassay for the quantitative determination of serotonin in urine and serum, to evaluate serotonin homeostasis. The determination of serotonin in urine is helpful for the determination of neurostress.

Contents of Kit

1. WASH-CONC 50x Wash Buffer Concentrate - concentrated 50x
Content: Buffer with a non-ionic detergent and physiological pH
Volume: 1 x 20 ml/vial, purple cap

2. CONJUGATE Antibody Conjugate – ready to use
Content: Goat anti-rabbit immunglobulins conjugated with peroxidase
Volume: 1 x 12 ml/vial
Description: Species is goat, red cap


3. SUBSTRATE Substrate – ready to use
Content: Chromogenic substrate containing 3,3',5,5'-tetramethylbenzidine, substrate buffer and hydrogen peroxide
Volume: 1 x 12 ml/vial, black cap


4. STOP-SOLN Stop Solution – ready to use
Content: 0.25 M sulfuric acid
Volume: 1 x 12 ml/vial, grey cap

5. Ш SER 5-HIAA Serotonin Microtiter Strips – ready to use
Content: 1 x 96 well (12x8) antibody precoated microwell plate in a resealable white pouch with desiccant

6. ACYL-REAG Acylation Reagent – ready to use
Content: Acylation reagent in DMSO
Volume: 2 x 3 ml/vial, white cap


7. SER-AS Serotonin Antiserum – ready to use
Content: Rabbit anti-Serotonin antibody, blue coloured
Volume: 1 x 6 ml/vial, blue cap
Description: Species is rabbit


8. ACYL-BUFF Acylation Buffer – ready to use
Content: TRIS buffer with non-mercury preservative
Volume: 1 x 55 ml/vial, grey cap

9. Calibration and Controls
Standards and Controls – ready to use
STANDARD A white 0 ng/ml, 0 nmol/l. 4 ml
STANDARD B yellow 15 ng/ml, 85 nmol/l. 4 ml
STANDARD C orange 50 ng/ml, 284 nmol/l. 4 ml
STANDARD D blue 150 ng/ml, 851 nmol/l. 4 ml
STANDARD E grey 500 ng/ml, ng/ml, 2,840 nmol/l. 4 ml
STANDARD F black 2,500 ng/ml, 14,175 nmol/l. 4 ml
CONTROL 1 green Refer to COA for expected value and acceptable range! 4 ml
CONTROL 2 red Refer to COA for expected value and acceptable range! 4 ml
Conversion: serotonin [ng/ml] x 5.67 = serotonin [nmol/l]
Content: TRIS buffer with non-mercury preservatives, spiked with defined quantity of serotonin

Storage

Store kit and reagents at 2 – 8°C until expiration date. Do not use components beyond the expiry date indicated on the kit labels. Once opened, the reagents are stable for 2 months when stored at 2 – 8°C. Once the resealable pouch has been opened, care should be taken to close it tightly again including the desiccant.

Performance Characteristics

Analytical Sensitivity was determined according to the CLSI Standard EP17-A2 Vol. 32 No. 8.
For the determination of the analytical sensitivity, 5 blank samples and 5 low level samples in 2 kit lots in 4 replicates per sample were determined. This resulted in 60 results blank and 60 results low level per lot.



The method comparison was conducted according to the CLSI standard CLSI EP09c 3rd ed.


The precision of the intra- and inter-assay variation was investigated by determining the concentration of 6 serum samples and 6 urine samples in two runs per day in each 2 replicates over 20 days (according to the CLSI standard EP05-A3 Vol. 34 No.13).

General Description

Serotonin (5-hydroxytryptamine) is an intermediate product of tryptophan metabolism, a wellstudied neurotransmitter, and may also act as a peripheral hormone. Synthesis occurs mainly in enterochromaffin cells (ec-cells) of the gastrointestinal tract and in neurons. It is present in high concentrations in ec-cells of the intestine, serotonergic neurons of the brain, and platelets. Serotonin is mainly degraded to 5-hydroxyindole acetic acid (5-HIAA) or melatonin and can be excreted in the urine. In the bloodstream, the vast majority of serotonin is found in platelets and can be readily detected in serum. Altered serotonin levels in serum and/or urine can indicate both physical and psychological dysfunction. Serotonin balance may be impaired in serum and/or urine in a variety of conditions. For example, decreased serotonin levels have been demonstrated in depression, anxiety, and even pain sensitivity compared to unaffected subjects. Increased serotonin levels, on the other hand, have been reported in patients with serotonin-secreting neuroendocrine tumors, also called carcinoid tumors, or hepatocellular carcinomas.

Standard Curve

Example do not use for calculation!

General Notes

Enzyme Immunoassay for the quantitative determination of serotonin in urine and serum, to evaluate serotonin homeostasis. The determination of serotonin in urine is helpful for the determination of neurostress.