RayBio® Human SPHK1 ELISA Kit for Cell Culture Supernatants, Plasma, and Serum samples.
Lead time: Typically ships within 1-2 business days. No Friday shipments.
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Product Description
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Estimated Lead Time | 1-2 business days |
| Species | Human |
| Sensitivity | 1.43 ng/ml Need more sensitivity? Check out the new NexaTag® kit for this target. Still not enough? Then your answer is our Ultrasensitive Biomarker Testing Service powered by Simoa™ technology. |
| Detection Range | 1.43 ng/ml - 400 ng/ml |
| Specificity | This ELISA kit shows no cross-reactivity with the following cytokines tested: human ADAMTSL-1/Punctin-1, AMSH/STAMBP, Annexin V, BATF3, Bora, Cadherin-17, Caveolin-2, CD2, CD200 R1, CHST3, COMT, Cystatin SA, Dopamine beta-Hydroxylase, Desmin, Exostosin-like 3, Ficolin-1, FosB/GOS3, FRS2, GATA-5, GFAP, GLI-3, HepaCAM, ARNT/HIF-1 beta, 17 betaHSD1/HSD17B1, IDO, Kallikrein 1, Kell, MDL-1/CLEC5A, NPDC-1, Numb, Olig2, p63/TP73L, Pax3, Semaphorin 4D, TAZ/WWTR1, TC-PTP, Transglutaminase 3/TGM3, TPST2, TREML1/TLT-1. |
| Compatible Sample Types | Cell Culture Supernatants, Plasma, Serum |
| Recommended Dilution (Serum/Plasma) | 2 fold |
| Accession Number | Q9NYA1 |
| Gene Id | 8877 |
| Gene Symbols | SPHK1, SPHK, SPK |
| Protein Name / Synonyms | Sphingosine kinase 1 (SK 1) (SPK 1) (EC 2.7.1.91) |
| Quantitative/Semi-Quantitative | Quantitative |
| Solid Support | 96-well Microplate |
| Design Principle | Sandwich-based |
| Method Of Detection | Colorimetric |
| Research Area | Angiogenesis, Cancer, Inflammation |
| Shipping Type | Blue ice |
| Storage | ≤-20°C |
Product Features
Additional Notes
- Strip plates and additional reagents allow for use in multiple experiments
- Quantitative protein detection
- Establishes normal range
- The best products for confirmation of antibody array data
Standard Curves
Spiking & Recovery Results
| Sample Type | Average % Recovery | Range (%) |
|---|---|---|
| Serum | 90.39 | 73-131 |
| Plasma | 98.92 | 72-127 |
| Cell culture media | 110.6 | 84-133 |
Linearity Results
| Sample Type | Serum | Plasma | Cell Culture Media | |
|---|---|---|---|---|
| 1:2 | Average % of Expected | 114.9 | 125.7 | 119.7 |
| Range (%) | 110-120 | 118-134 | 118-121 | |
| 1:4 | Average % of Expected | 125.1 | 121.5 | 112.5 |
| Range (%) | 111-139 | 116-125 | 110-115 | |
Application Notes
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Stop Solution
- Assay Diluent(s)
- Lyophilized Standard
- Biotinylated Detection Antibody
- Streptavidin-Conjugated HRP
- TMB One-Step Substrate
Other Materials Required
- Distilled or deionized water
- Precision pipettes to deliver 2 µl to 1 µl volumes
- Adjustable 1-25 µl pipettes for reagent preparation
- 100 µl and 1 liter graduated cylinders
- Tubes to prepare standard and sample dilutions
- Absorbent paper
- Microplate reader capable of measuring absorbance at 450nm
- Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
- Prepare all reagents, samples and standards as instructed in the manual.
- Add 100 µl of standard or sample to each well.
- Incubate 2.5 h at RT or O/N at 4°C.
- Add 100 µl of prepared biotin antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared Streptavidin solution to each well.
- Incubate 45 min at RT.
- Add 100 µl of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl of Stop Solution to each well.
- Read at 450 nm immediately.
Storage/Stability
The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
