Added to cart successfully !

QuicKey Human ADP/Acrp30(Adiponectin) ELISA Kit

Size:  
Qty:
- +
Price:   $390
Cat Number:  E-TSEL-H0020
Lead Time:   7~10 days

Reactivity:  Human

Detection Range:  0.39~25 ng/mL

Sensitivity:  0.18 ng/mL

Add to cart

Product Details

QuicKey Series

Get more sensitive and precise results with saving at least 1h comparing to traditional ELISA Kits. The new developed technology in house will help to accelerate your science research in a more efficient way.

Properties

Assay type Sandwich-ELISA
Format 96T/48T
Assay time 2.5 h
Reactivity Human
Detection method Colormetric
Colormetric 0.39-25 ng/mL
Sensitivity 0.18 ng/mL
Sample volume 50μL/well
Sample type serum, plasma
Specificity This kit recognizes Human ADP/Acrp30 in samples. No significant cross-reactivity or interference between Human ADP/Acrp30 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 Human ADP/Acrp30 concentrations in serum, plasma. Please consult technical support for the applicability if other biological fluids need to be tested.

Test principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human ADP/Acrp30. Samples (or Standards) and biotinylated detection antibody specific for Human ADP/Acrp30 are added to the micro ELISA plate wells. Human ADP/Acrp30 would combine with the specific antibody. Then Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human ADP/Acrp30, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 ± 2 nm. The OD value is proportional to the concentration of Human ADP/Acrp30. You can calculate the concentration of Human ADP/Acrp30 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for six months. After test, the unused wells and reagents should be stored according to the table below. (The specific components might be slightly different in delivered kits, please refer to the manual for more information.)

Item Specifications Storage conditions after test
Micro ELISA Plate (Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips
2-8℃, 1 month
Reference Standard 96T: 2 vials
48T: 1 vial
Discard unused reconstituted standard and dilutions
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8℃
Biotinylated Detection Ab Working Solution 1 vial, 6 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
2-8℃ (Protect from light)
Substrate Reagent 1 vial, 10 mL
Stop Solution 1 vial, 10 mL 2-8℃
Plate Sealer 5 pieces
Product Description 1 copy
Certificate of Analysis 1 copy

Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution. The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

Other supplies required

1. Microplate reader with 450nm wavelength filter

2. High-precision transfer pipette, EP tubes and disposable pipette tips

3. Incubator capable of maintaining 37℃

4. Deionized or distilled water

5. Absorbent paper

6. Loading slot for Wash Buffer

Technical Data

Sample values

sandwich-Ab-ELISA-Elabscience

Typical Standard Curve

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration(ng/mL) 25 12.5 6.25 3.13 1.56 0.78 0.39 0
OD 2.384 1.519 0.888 0.547 0.36 0.208 0.143 0.070
Corrected OD 2.314 1.449 0.817 0.477 0.29 0.138 0.073 -
sandwich-Ab-ELISA-Elabscience

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human ADP/Acrp30 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human ADP/Acrp30 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(ng/mL) 1.09 2.57 12.28 1.24 2.82 11.20
Standard deviation 0.06 0.12 0.49 0.07 0.14 0.55
CV (%) 5.50 4.67 3.99 5.65 4.96 4.91

Recovery

The recovery of Human ADP/Acrp30 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) 97-109 102
EDTA plasma(n=8) 87-99 92

Linearity

Samples were spiked with high concentrations of Human ADP/Acrp30 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum(n=4) EDTA plasma (n=4)
1:2 Range (%) 98-108 91-104
Average (%) 102 96
1:4 Range (%) 90-102 99-108
Average (%) 95 105
1:8 Range (%) 89-101 96-105
Average (%) 97 101
1:16 Range (%) 91-102 97-106
Average (%) 94 102

Target information

Background

Adiponectin(Acrp30/ADP), is a hormone exclusively secreted by adipocytes that regulates energy homeostasis, and glucose and lipid metabolism[1]. In human, it is a 244 amino acid, 30kDa, protein encoded by gene apM1, that shares structural homology with collagen Ⅷ and Ⅹ and complement factor C1q[2]. Adiponectin assembles into different complexes including trimers (low molecular weight), hexamers (middle molecular weight), and higher order oligomeric structures (high molecular weight) that may affect biological activity[3]. Adiponectin is an adipocyte-derived protein with wide ranging paracrine and endocrine effects on metabolism and inflammation. It promotes adipocyte differentiation, fatty acid catabolism, and insulin sensitivity. Research has shown that adiponectin is associated with many kinds of diseases, such as obesity, insulin resistance, type 2 diabetes, hyperlipidemia, hypertension, atherogenesis and renal disease[2].
1. Hu E, Liang P, Spiegelman B M. AdipoQ is a novel adipose-specific gene dysregulated in obesity[J]. Journal of biological chemistry, 1996, 271(18): 10697-10703.
2. J Diez J, Iglesias P. The role of the novel adipocyte-derived protein adiponectin in human disease: an update[J]. Mini reviews in medicinal chemistry, 2010, 10(9): 856-869.
3. Pajvani U B, Du X, Combs T P, et al. Structure-function studies of the adipocyte-secreted hormone Acrp30/adiponectin implications for metabolic regulation and bioactivity[J]. Journal of Biological Chemistry, 2003, 278(11): 9073-9085.

Research area

Database Links

SwissProt Q15848 Human
Entrez Gene 9370 Human

Synonyms

GBP28, ACDC, APM1, ADPN, AdipoQ, ADIPQTL1

Documents

Citations

Please kindly contact us if you have published a paper by citing any products from Elabscience, we will get back to you with the Elabscience Publication Reward ASAP.

Review more published papers in our Publication Library

Assay procedures

1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 90 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
If you have any question about the product, please feel free to contact us or leave a message