General species Sphingosine-1-Phosphate (S1P) (Competitive EIA) ELISA Kit

Mfr.No.

CEG031Ge

Sensitivity

4.62ng/mL

Detection Range

12.35-1,000ng/mL

Reactivity

General

Assay Duration

Assay Type

Competitive Inhibition

Description

This assay has high sensitivity and excellent specificity for detection of Sphingosine-1-Phosphate (S1P).
No significant cross-reactivity or interference between Sphingosine-1-Phosphate (S1P) and analogues was observed.

Detection Method

Colormetric

Kit Components

Reagents	Quantity	Reagents	Quantity
Pre-coated, ready to use 96-well strip plate	1	Plate sealer for 96 wells	4
Standard	2	Standard Diluent	1×20mL
Detection Reagent A	1×120µL	Assay Diluent A	1×12mL
Detection Reagent B	1×120µL	Assay Diluent B	1×12mL
TMB Substrate	1×9mL	Stop Solution	1×6mL
Wash Buffer (30 × concentrate)	1×20mL	Instruction manual	1

Recovery

Matrices listed below were spiked with certain level of Sphingosine-1-Phosphate (S1P) and the recovery rates were calculated by comparing the measured value to the expected amount of Sphingosine-1-Phosphate (S1P) in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	94-105	98
EDTA plasma(n=5)	81-98	86
heparin plasma(n=5)	80-103	82

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Sphingosine-1-Phosphate (S1P) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1:2	1:4	1:8	1:16
serum(n=5)	89-101%	94-103%	82-99%	82-104%
EDTA plasma(n=5)	81-94%	90-98%	78-96%	84-97%
heparin plasma(n=5)	98-105%	78-105%	86-95%	78-96%

Features

Detects small molecule

Species

General species

Sample Type

Cell lysates, Cell culture supernates, Serum, Tissue homogenates, Plasma, Other biological fluids

Precautions

The stability of 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.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Sphingosine-1-Phosphate (S1P) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Sphingosine-1-Phosphate (S1P) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: C<10%
Inter-Assay: C<12%

Notes

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to S1P has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled S1P analogues and unlabeled antigen (Standards or samples) with the pre-coated antibody. After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of S1P in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of S1P in the sample.

Usage Notes

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
And then add 50µL prepared Detection Reagent A immediately.
Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

Applications	ELISA

Storage	4°C for 1 month/ -20°C for long-term(within shelf life)
Shelf Life	one year

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SKU	Supplier No.	Size	Your price
BHE10107128	CEG031Ge	96T*1	$940	Add to Cart
BHE10107128	CEG031Ge	96T*5	$4230	Add to Cart
BHE10107128	CEG031Ge	96T*10	$7990	Add to Cart

ELISA Kit for Sphingosine-1-Phosphate (S1P) (General species)

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