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IP3(Inositol Triphosphate) ELISA Kit

Cat.No.:E-EL-0059
Reactivity: Universal

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	15.63-1000 pg/mL
Sensitivity	9.38 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes IP3 in samples. No significant cross-reactivity or interference between IP3 and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of IP3 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with IP3. During the reaction, IP3 in the sample or standard competes with a fixed amount of IP3 on the solid phase supporter for sites on the Biotinylated Detection Ab specific to IP3. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of IP3 in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8℃
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level IP3 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level IP3 were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	54.40	130.40	426.20	58.40	132.30	424.50
Standard deviation	2.70	5.20	16.20	4.00	7.70	14.90
CV (%)	4.96	3.99	3.80	6.85	5.82	3.51

Recovery

The recovery of IP3 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	88-100	93
EDTA plasma(n=8)	88-98	93
Cell culture media(n=8)	85-101	92

Linearity

Samples were spiked with high concentrations of IP3 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Synonyms	Inositol trisphosphate, inositol 1, 4, 5-trisphosphate, triphosphoinositol, InsP3, Ins3P, IP3
Research Area	Immunology, Cancer

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results

Citations

Molecular Cancer (2019) IF: 10.679
Epigenetically upregulated oncoprotein PLCE1 drives esophageal carcinoma angiogenesis and proliferation via activating the PI-PLCε-NF-κB signaling pathway and VEGF-C/ Bcl-2 expression

DOI: 10.1186/s12943-018-0930-x

Sample: Esophageal squamous cell carcinoma (ESCC) cells
Communications Biology (2021) IF: 6.268
Iron induces two distinct Ca2+ signalling cascades in astrocytes

DOI: 10.1038/s42003-021-02060-x

Sample: Cell culture supernatant
Nutrients (2022) IF: 6.706
Dihydromyricetin Enhances Exercise-Induced GLP-1 Elevation through Stimulating cAMP and Inhibiting DPP-4

DOI: 10.3390/nu14214583
ENDOCRINOLOGY (2023) IF: 5.051
Aspirin suppresses hepatic glucagon signaling through decreasing production of thromboxane A2

DOI: 10.1210/endocr/bqac217

Sample: Hepatocyte
Oxidative Medicine and Cellular Longevity (2020) IF: 5.076
The Degree of Helicobacter pylori Infection Affects the State of Macrophage Polarization through Crosstalk between ROS and HIF-1α

DOI: 10.1155/2020/5281795

Sample: Cell culture supernatant
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS (2022) IF: 4.117
A2A R mediated modulation in IP3 levels altering the [Ca2+]i through cAMP-dependent PKA signalling pathway

DOI: 10.1016/j.bbagen.2022.130242

PMID: 36084905
Sample: HEK 293 cell
FEBS Journal (2019) IF: 4.739
Impaired calcium homeostasis via advanced glycation end products promotes apoptosis through endoplasmic reticulum stress in human nucleus pulposus cells and exacerbates intervertebral disc degeneration in rats

DOI: 10.1111/febs.14972

PMID: 31230413
Sample: Cell culture supernatant,Tissue homogenate
EXPERIMENTAL CELL RESEARCH (2022) IF: 3.905
A new missense mutation of calcium sensing receptor with isoleucine replaced by serine at codon 857 leading to type V Bartter syndrome

DOI: 10.1016/j.yexcr.2022.113080

PMID: 35192837
Sample: Urine stem cell
Scientific Reports (2020) IF: 3.998
From extraocular photoreception to pigment movement regulation: a new control mechanism of the lanternshark luminescence

DOI: 10.1038/s41598-020-67287-w

PMID: 32576969
Sample: Tissue homogenate

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