Name: Estradiol ELISA Kit
Brand: Hormone & Small Molecule ELISA Kits
SKU: UNFI0011
Availability: OutOfStock

Key Features

	Save Time	Pre-coated 96 well plate
	Quick Start	Kit includes all necessary reagents
	Publication Ready	Reproducible and reliable results

Overview


Product Name:	E2 (Estradiol) ELISA Kit
Product Code:	UNFI0011
Size:	96 Assays
Target:	E2
Alias:	E2, Estradiol
Reactivity:	Universal
Detection Method:	Competitive ELISA, Coated with Antibody
Sensitivity:	7.5pg/ml
Range:	12.5-800pg/ml
Storage:	4°C for 6 months
Note:	For Research Use Only

Additional Information

Recovery:

Matrices listed below were spiked with certain level of E2 and the recovery rates were calculated by comparing the measured value to the expected amount of E2 in samples.


Matrix	Recovery Range (%)	Average (%)
serum(n=5)	87-103	95
EDTA plasma(n=5)	91-105	98
UFH plasma(n=5)	88-104	99

Linearity:

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of E2 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.


Sample	1:2	1:4	1:8
serum(n=5)	85-101%	97-103%	88-100%
EDTA plasma(n=5)	85-101%	84-101%	85-100%
UFH plasma(n=5)	81-96%	88-98%	82-95%

Intra-Assay:

CV <8%

Inter-Assay:

CV <10%

Protocol

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Step	Procedure
1.	Set standard, test sample and control (zero) wells on the pre-coatedplate respectively, and then, record their positions. It isrecommended to measure each standard and sample in duplicate. Washplate 2 times before adding standard, sample and control (zero) wells!
2.	Add Sample and Biotin-detection antibody: Add 50µL of Standard, Blank or Sample per well. The blankwell is added with Sample Dilution Buffer. Immediately add 50 µL of biotin-labelled antibody workingsolution to each well. Cover with the plate sealer provided. Gently tap the plate to ensure thoroughmixing. Incubate for 45 minutes at 37°C. (Solutions are added to the bottom of micro-ELISA platewell, avoid touching plate walls and foaming).
3.	Wash: Aspirate each well and wash, repeating the process three timesWash by filling each well with Wash Buffer (approximately 350µL)using a squirt bottle, multi-channel pipette, manifold dispenser orautomated washer. Complete removal of liquid at each step is essentialto good performance. After the last wash, remove any remaining WashBuffer by aspirating or decanting. Invert the plate and pat it againstthick clean absorbent paper.
4.	HRP-Streptavidin Conjugate(SABC): Add 100µL of SABC workingsolution to each well. Cover with a new Plate sealer. Incubate for30minutes at 37°C.
5.	Wash: Repeat the aspiration/wash process for five times.
6.	TMB Substrate: Add 90µL of TMB Substrate to each well. Coverwith a new Plate sealer. Incubate for about 10-20 minutes at 37°C.Protect from light. The reaction time can be shortened or extendedaccording to the actual color change, but not more than 30minutes.When apparent gradient appeared in standard wells, you can terminatethe reaction.
7.	Stop: Add 50µL of Stop Solution to each well. Color turn toyellow immediately. The adding order of stop solution should be as thesame as the substrate solution.
8.	OD Measurement: Determine the optical density (OD Value) of each wellat once, using a microplate reader set to 450 nm. You should open themicroplate reader ahead, preheat the instrument, and set the testing parameters.

Sample Preparation

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type

Protocol

Serum

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