Multiplex Assay Kit for Sphingosine-1-Phosphate (S1P) ,etc. by CBA (Cytometric Bead Array)
Overview
Properties
- Product No.CMG031Ge
- Organism SpeciesPan-species (General) Same name, Different species.
- ApplicationsFLIA Kit for Antigen Detection.
Research use only - DownloadInstruction Manual
- Category
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Recovery
Matrices listed below were spiked with certain level of Sphingosine-1-Phosphate (S1P) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Sphingosine-1-Phosphate (S1P) ,etc. by CBA (Cytometric Bead Array) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 82-102 | 90 |
| EDTA plasma(n=5) | 89-96 | 92 |
| heparin plasma(n=5) | 79-97 | 82 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Sphingosine-1-Phosphate (S1P) ,etc. by CBA (Cytometric Bead Array) 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) ,etc. by CBA (Cytometric Bead Array) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Sphingosine-1-Phosphate (S1P) ,etc. by CBA (Cytometric Bead Array) 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) | 84-94% | 97-105% | 87-95% | 96-103% |
| EDTA plasma(n=5) | 80-101% | 91-103% | 96-104% | 90-99% |
| heparin plasma(n=5) | 95-103% | 87-95% | 93-103% | 96-105% |
Stability
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.
Reagents and materials provided
| Reagents | Quantity | Reagents | Quantity |
| 96-well plate | 1 | Plate sealer for 96 wells | 4 |
| Pre-Mixed Standard | 2 | Standard Diluent | 1×20mL |
| Pre-Mixed Magnetic beads (22#:S1P) | 1 | Analysis buffer | 1×20mL |
| Pre-Mixed Detection Reagent A | 1×120μL | Assay Diluent A | 1×12mL |
| Detection Reagent B (PE-SA) | 1×120μL | Assay Diluent B | 1×12mL |
| Sheath Fluid | 1×10mL | Wash Buffer (30 × concentrate) | 1×20mL |
| Instruction manual | 1 |
Assay procedure summary
1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 50μL standard or sample to each well, add 50μL prepared Detection Reagent A, add 10μL magnetic beads, and incubate 90 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
5. Wash plate once on magnetic frame;
6. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.
Test principle
Anti-Sphingosine-1-Phosphate (S1P) ,etc. by CBA (Cytometric Bead Array) antibody was coated on the surface of magnetic beads. A competitive inhibition reaction is launched between biotin labeled Sphingosine-1-Phosphate (S1P) ,etc. by CBA (Cytometric Bead Array) and unlabeled Sphingosine-1-Phosphate (S1P) ,etc. by CBA (Cytometric Bead Array) (Standards or samples) with the pre-coated antibody specific to Sphingosine-1-Phosphate (S1P) ,etc. by CBA (Cytometric Bead Array). PE-labeled avidin was added. The amount of bound PE conjugate is reverse proportional to the concentration of Sphingosine-1-Phosphate (S1P) ,etc. by CBA (Cytometric Bead Array) in the sample, the intensity of MFI developed is reverse proportional to the concentration of Sphingosine-1-Phosphate (S1P) ,etc. by CBA (Cytometric Bead Array) in the sample.
Giveaways
Increment services
Citations
- Quercetin ameliorates pulmonary fibrosis by inhibiting SphK1/S1P signalingPubmed:29940125
- Nppb Neurons Are Sensors of Mast Cell-Induced ItchPubmed: 30917312
- SPNS 2 promotes the malignancy of colorectal cancer cells via regulating Akt and ERK pathwayPubmed: 31206801
- Role of miR-506 in ulcerative colitis associated with primary sclerosing cholangitis33980925
- The S1PR1 Agonist SEW2871 promotes the Survival of Skin Flap34310896
- Berberine ameliorates erectile dysfunction in rats with streptozotocin‐induced diabetes mellitus through the attenuation of apoptosis by inhibiting the SPHK1/S1P …34674380
- α-Ketoglutarate Inhibits Thrombosis and Inflammation by Prolyl Hydroxylase-2 Mediated Inactivation of Phospho-Akt34740102
- Implication of Sphingolipid Metabolism Gene Dysregulation and Cardiac Sphingosine-1-Phosphate Accumulation in Heart FailurePubmed:35052814
- Sphk2 deletion is involved in structural abnormalities and Th17 response but does not aggravate colon inflammation induced by sub-chronic stressPubmed:35260749
- Anti-inflammatory activity of non-selective PDE inhibitor aminophylline on the lung tissue and respiratory parameters in animal model of ARDS
- Network Pharmacology Study on the Diuretic Effect of Plantaginis Semen and its Anti-Hypertensive Mechanism
