Hormone & Small Molecule ELISA Kits

HA / Hyaluronic Acid ELISA Kit

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SKU:
UNFI0033
Product Type:
ELISA Kit
Size:
96 Assays
Sensitivity:
0.938ng/ml
Range:
1.563-100ng/ml
ELISA Type:
Competitive
Synonyms:
HA, Hyaluronic Acid
Reactivity:
Universal

Description

HA / Hyaluronic Acid ELISA Kit

system_update_alt Datasheet system_update_alt MSDS

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:

HA (Hyaluronic Acid) ELISA Kit

Product Code:

UNFI0033

Size:

96 Assays

Target:

HA

Alias:

HA, Hyaluronic Acid

Reactivity:

Universal

Detection Method:

Competitive ELISA, Coated with Antibody

Sensitivity:

0.938ng/ml

Range:

1.563-100ng/ml

Storage:

4°C for 6 months

Note:

For Research Use Only

Additional Information

Recovery

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

Matrix

Recovery Range (%)

Average (%)

serum(n=5)

85-103

95

EDTA plasma(n=5)

93-104

100

UFH plasma(n=5)

91-103

96

Linearity:

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HA 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)

86-105%

87-99%

88-101%

EDTA plasma(n=5)

83-97%

86-100%

82-97%

UFH plasma(n=5)

80-93%

93-99%

84-98%

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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