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5-Hydroxytryptamine (5HT) ELISA Kit


Species Reactivity

General

UniProt

Abbreviation

5HT

Alternative Names

N/A

Range

3.12-200 ng/mL

Sensitivity

1 ng/mL

Sample Type

Serum, Plasma, Other biological fluids

Detection Method

Sandwich

Analysis Method

Quantitive

Assay Duration

1-4.5h

Sample Volume

1-200 μL

Detection Wavelengt

450 nm

Cat

AE64465GE

Size

96T 48T 96T*5 96T*50

Price

Inquire

Availability

3-5 working days

Manual

Inquire
Test principle:
This assay employs a two-site sandwich ELISA to quantitate 5HT in samples. An antibody specific for 5HT has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any5HT present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for 5HT is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of 5HT bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:
Serotonin also called 5-hydroxytryptamine, is primarily found in the gastrointestinal (GI) tract and central nervous system (CNS) of animals. Approximately 80 percent of the human body's total serotonin is located in the enterochromaffin cells in the gut, where it is used to regulate intestinal movements. The remainder is synthesized in serotonergic neurons in the CNS where it has various functions, including the regulation of mood, appetite, sleep, muscle contraction, and some cognitive functions including memory and learning; and in blood platelets where it helps to regulate hemostasis and blood clotting. In addition to animals, serotonin is also found in fungi and plants. 5-hydroxytryptamine(5-HT) is an important neurotransmitter made by neurons in the central nervous system. These functions are mediated by a large number of receptors.After it is released by a neuron, 5-HT activates receptors located on nearby neurons.
Components:

Reagents

Quantity

Reagents

Quantity

Assay plate (96 Wells)

1

Instruction manual

1

Standard (lyophilized)

2

Sample Diluent

1 x 20 mL

Biotin-Conjugate (concentrate 100 x)

1 x 120 μL

Biotin-Conjugate Diluent

1 x 20 mL

Streptavidin-HRP  (concentrate 100 x)

1 x 120 μL

Streptavidin-HRP Diluent   

1 x 20 mL

Wash Buffer (concentrate 25 x)

1 x 20 mL

Substrate Solution

1 x 12 mL

Stop Solution

1 x 10 mL

Adhesive Films

4

Specificity:
This assay has high sensitivity and excellent specificity for detection of General5HT. No significant cross-reactivity or interference between General5HT and analogues was observed.
Recovery:
Matrices listed below were spiked with certain level of recombinant General5HT and the recovery rates were calculated by comparing the measured value to the expected amount of General5HT in samples.

Sample Type

Number

Recovery range (%)

Average(%)

Serum

10

90-101

96

EDTA plasma

10

89-97

93

Heparin plasma

10

91-99

95

Precision:
Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
Linearity:
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of General5HT and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample Type

1:2

1:4

1:8

1:16

Serum

78-89%

81-99%

92-103%

95-105%

EDTA plasma

91-101%

90-98%

93-101%

91-98%

Heparin plasma

92-103%

93-102%

92-99%

91-101%

Stability:
The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
Sample collection and storage:
Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Kits storage instructions:
Store at 2-8°C. Please refer to Instruction Manual.