亞精胺 (SMD) ELISA 試劑盒| Spermidine (SMD) ELISA Kit abbexa貨號abx585001－太鼎生物科技/太鼎食安科技

亞精胺 (SMD) ELISA 試劑盒| Spermidine (SMD) ELISA Kit貨號abx585001

偵測範圍0.78 ng/ml - 200 ng/ml

亞精胺 (SMD) ELISA 試劑盒是用於體外定量測定血清、血漿、組織勻漿、細胞裂解物、細胞培養上清液和其他生物體液中亞精胺 (SMD) 濃度的 ELISA 試劑盒。Spermidine (SMD) ELISA Kit is an ELISA Kit for the in vitro quantitative measurement of Spermidine (SMD) concentrations in serum, plasma, tissue homogenates, cell lysates, cell culture supernatants and other biological fluids.

產品介紹:

亞精胺(Spermidine)是一種多胺化合物 (polyamine compound , C7H19N3)，存在於核醣體(ribosomes)和活組織(living tissues)中，在生物體(organisms)內具有多種代謝功能(metabolic functions)。它最初是從精液(semen)中分離出來的。多胺(Polyamines)，例如亞精胺(spermidine)，是聚陽離子脂肪胺(polycationic aliphatic amines)並且是多功能(multifunctional)的。它們在細胞存活中起著至關重要的作用。亞精胺合酶 (Spermidine synthase ,SPDS) 催化腐胺(putrescine)形成亞精胺(spermidine)。亞精胺(spermidine)是進一步多胺(polyamines)的前體，例如精胺(spermine)及其結構異構體熱精胺(isomer thermospermine)。亞精胺(Spermidine synchronizes)同步一系列生物過程（例如 Ca2+、Na+、K+ -ATP 酶），從而維持膜電位並控制細胞內 pH 和體積。亞精胺調節生物過程，例如穀氨酸能 N 甲基-d-天冬氨酸受體（glutamatergic Nmethyl- d-aspartate receptor ,NMDA 受體）引起的 Ca2+ 流入，這與一氧化氮合酶 (nitric oxide synthase ,NOS) 和 cGMP/PKG 通路激活以及大腦皮層突觸體(cerebral cortex synaptosomes) Na+、K+-ATP 酶活性降低有關。

In troduction	Spermidine is a polyamine compound (C7H19N3) found in ribosomes and living tissues, and having various metabolic functions within organisms. It was originally isolated from semen. Polyamines, such as spermidine, are polycationic aliphatic amines and are multifunctional. They serve vital roles in cell survival. Spermidine synthase (SPDS) catalyzes the formation of spermidine from putrescine. Spermidine is a precursor to further polyamines, such as spermine and its structural isomer thermospermine. Spermidine synchronizes an array of biological processes (such as Ca2+, Na+, K+ -ATPase) thus maintaining membrane potential and controlling intracellular pH and volume. Spermidine regulates biological processes, such as Ca2+ influx by glutamatergic Nmethyl- d-aspartate receptor (NMDA receptor), which has been associated with nitric oxide synthase (NOS) and cGMP/PKG pathway activation and a decrease of Na+,K+-ATPase activity in cerebral cortex synaptosomes.
Target	Spermidine (SMD)
Reactivity	General (All species)
Tested Applications	ELISA
Recommended dilutions	Optimal dilutions/concentrations should be determined by the end user.
Storage	Shipped at 4 °C. Upon receipt, store the kit according to the storage instruction in the kit's manual.
Validity	The validity for this kit is 6 months.
Stability	The stability of the kit is determined by the rate of activity loss. The loss rate is less than 5% within the expiration date under appropriate storage conditions. To minimize performance fluctuations, operation procedures and lab conditions should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same user throughout.
Test Range	0.78 ng/ml - 200 ng/ml
Sensitivity	< 0.29 ng/ml
Standard Form	Lyophilized
Detection Method	Colorimetric
Assay Type	Competitive
Assay Data	Quantitative
Sample Type	Serum, plasma, tissue homogenates, cell lysates, cell culture supernatants and other biological fluids.
Target Type	Antigen
Assay Principle	This kit is based on competitive enzyme-linked immuno-sorbent assay technology. An antibody is pre-coated onto a 96-well plate. Standards, test samples, and biotin-conjugated reagent are added to the wells and incubated. A competitive inhibition reaction takes place between the biotin-labelled SMD and the unlabelled- SMD on the pre-coated antibody. The HRP-conjugated reagent is then added, and the whole plate is incubated. Unbound conjugates are removed using wash buffer at each stage. TMB substrate is used to quantify the HRP enzymatic reaction. After TMB substrate is added, only wells that contain sufficient SMD will produce a blue coloured product, which then changes to yellow after adding the acidic stop solution. The intensity of the color yellow is inversely proportional to the SMD amount bound on the plate. The OD is measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of SMD can be calculated.
Kit Components	The kit components listed are for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product. Pre-coated 96-Well Microplate Standard Standard Diluent Buffer Wash Buffer Detection Reagent A Detection Reagent B Diluent A Diluent B TMB Substrate Stop Solution Plate Sealer
Material Required But Not Provided	37°C incubator Multi and single channel pipettes and sterile pipette tips Squirt bottle or automated microplate washer 1.5 ml tubes Distilled water Absorbent filter papers 100 ml and 1 liter graduated cylinders Microplate reader (wavelength: 450 nm) ELISA Shaker
Reagent Preparation	This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product. 1) Standard: Prepare the standard with the recommended volume of Standard Diluent Buffer, to make the standard solution. Then use the Standard Diluent buffer to carry out serial dilutions of the standard solution, as instructed in the Protocol. 2) Wash Buffer: Dilute the concentrated Wash Buffer with distilled water, as instructed in the Protocol. 3) Detection Reagent Preparation: Calculate the total volume of working solution required. Dilute Detection Reagent A and Detection Reagent B with Diluent A and Diluent B, respectively, at 1:100.
Assay Procedure	This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product. 1) Set standard, test samples and control wells. 2) Aliquot 50 µl of diluted standard into the standard wells. 3) Aliquot 50 µl of Standard Diluent buffer into the control (zero) well. 4) Aliquot 50 µl of diluted samples into the sample wells. 5) Immediately aliquot 50 µl of Detection Reagent A to each well. Incubate for 1 hr at 37 °C. 6) Wash 3 times. 7) Aliquot 100 µl of Detection Reagent B to each well. Incubate for 30 mins at 37 °C. 8) Wash 5 times. 9) Aliquot 90 µl of TMB Substrate to each well. Incubate for 10-20 mins at 37 °C. 10) Aliquot 50 µl of Stop Solution. 11) Measure the OD at 450 nm.
Protocol	This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product. Equilibrate the kit components and samples to room temperature (18 - 25 °C) before use. It is recommended to plot a standard curve for each test. 1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample at least in duplicate. 2. Add 50 µL of each standard, control and sample into the appropriate wells. 3. Remove the cover and discard the liquid. 4. Immediately aliquot 50 µl of Detection Reagent A working solution. Seal the plate with a cover and incubate for 1 h at 37°C. 5. Remove the cover and discard the solution. Wash the plate 3 times with 1X Wash Buffer. 6. Add 100 µL of Detection Reagent B working solution into each well, seal and incubate at 37°C for 30 min. 7. Discard the solution and wash the plate 5 times with wash buffer as explained in previous step. 8. Aliquot 90 µl of TMB Substrate into each well. Seal the plate with a cover and incubate at 37°C for 10-20 min. Avoid exposure to light. The incubation time is for reference use only, the optimal time should be determined by end user. Do not exceed 30 min. 9. Add 50 µL of Stop Solution to each well. Read at 450 nm immediately.
Results Calculation	This assay is competitive, therefore there is an inverse correlation between SMD concentration in the sample and the absorbance measured. Create a graph with the log of the standard concentration (y-axis) and average absorbance measured (x-axis). Apply a best fit trendline through the standard points. The SMD concentration of the samples can be interpolated from the standard curve.
Assay Precision	Intra-assay Precision (Precision within an assay): 3 samples with low, medium and high levels of Spermidine (SMD) were were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, medium and high levels of Spermidine (SMD) were tested on 3 different plates, 8 replicates in each plate. CV (%) = (Standard Deviation / mean) × 100 Intra-Assay: CV<10% Inter-Assay: CV<12%
Availability	Shipped within 1-2 weeks.
Note	This product is for research use only.The range and sensitivity is subject to change. Please contact us for the latest product information. For accurate results, sample concentrations must be diluted to mid-range of the kit. If you require a specific range, please contact us in advance or write your request in your order comments.Please note that our ELISA and CLIA kits are optimised for detection of native samples, rather than recombinant proteins. We are unable to guarantee detection of recombinant proteins, as they may have different sequences or tertiary structures to the native protein.
Standard	200 ng/ml