Mouse ADIPOR1 (Adiponectin Receptor 1) ELISA Kit (MOES01149)

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MOES01149
€549

Description

ELISA Kit Technical ManualMSDS

Mouse ADIPOR1 (Adiponectin Receptor 1) ELISA Kit

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse ADIPOR1 . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse ADIPOR1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse ADIPOR1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse ADIPOR1. The concentration of Mouse ADIPOR1 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
Detection MethodColormetric
Detection Range3.13-200 ng/mL
Sensitivity1.88 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Mouse ADIPOR1 in samples. No significant cross-reactivity or interference between Mouse ADIPOR1 and analogues was observed.

Typical Data

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)
O.D Average Corrected
2002.294
2.348
2.3212.244
1001.519
1.541
1.531.453
500.878
0.854
0.8660.789
250.399
0.433
0.4160.339
12.50.255
0.249
0.2520.175
6.250.173
0.165
0.1690.092
3.130.117
0.133
0.1250.048
00.076
0.078
0.077--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse ADIPOR1 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse ADIPOR1 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)10.3330.2985.8310.7631.4387.33
Standard deviation0.581.543.290.741.712.79
C V (%)5.615.083.836.885.443.19

Recovery

The recovery of Mouse ADIPOR1 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=5)94-110101
EDTA plasma (n=5)87-9994
Cell culture media (n=5)94-110102

Linearity

Samples were spiked with high concentrations of Mouse ADIPOR1 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)86-9783-9893-108
Average (%)919099
1:4Range (%)85-9984-9786-99
Average (%)928993
1:8Range (%)90-10787-9786-101
Average (%)989292
1:16Range (%)90-10282-9583-95
Average (%)968790

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Mouse ADIPOR1 (Adiponectin Receptor 1) ELISA Kit (MOES01149) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
    7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Mouse ADIPOR1 (Adiponectin Receptor 1) ELISA Kit (MOES01149) Protein Information

UniProt Protein Function:ADIPOR1: Receptor for globular and full-length adiponectin (APM1), an essential hormone secreted by adipocytes that acts as an antidiabetic. Probably involved in metabolic pathways that regulate lipid metabolism such as fatty acid oxidation. Mediates increased AMPK, PPARA ligand activity, fatty acid oxidation and glucose uptake by adiponectin. Has some high-affinity receptor for globular adiponectin but low-affinity receptor for full-length adiponectin. Belongs to the ADIPOR family.
UniProt Protein Details:

Protein type:Membrane protein, multi-pass; Membrane protein, integral

Cellular Component: integral to membrane; intrinsic to plasma membrane; membrane; plasma membrane

Molecular Function:adiponectin binding; identical protein binding; metal ion binding; protein binding; protein heterodimerization activity; protein kinase binding; receptor activity

Biological Process: adiponectin-mediated signaling pathway; fatty acid metabolic process; fatty acid oxidation; G-protein coupled receptor protein signaling pathway; glucose homeostasis; hormone-mediated signaling; leptin-mediated signaling pathway; lipid metabolic process; negative regulation of cell growth; positive regulation of insulin receptor signaling pathway; positive regulation of JAK-STAT cascade; regulation of lipid metabolic process

NCBI Summary:This gene encodes a receptor for the fat-derived hormone adiponectin. Binding of adiponectin to the encoded protein results in activation of an AMP-activated kinase signaling pathway which affects levels of fatty acid oxidation and insulin sensitivity. Homozygous knockout mice for this gene exhibit elevated plasma glucose and insulin levels as well as impaired glucose tolerance. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Apr 2015]
UniProt Code:Q91VH1
NCBI GenInfo Identifier:807045897
NCBI Gene ID:72674
NCBI Accession:NP_001292998.1
UniProt Secondary Accession:Q91VH1,Q9CZA0,
UniProt Related Accession:Q91VH1
Molecular Weight:42,366 Da
NCBI Full Name:adiponectin receptor protein 1
NCBI Synonym Full Names:adiponectin receptor 1
NCBI Official Symbol:Adipor1  
NCBI Official Synonym Symbols:Paqr1; ACDCR1; CGI-45; 2810031L11Rik  
NCBI Protein Information:adiponectin receptor protein 1
UniProt Protein Name:Adiponectin receptor protein 1
Protein Family:Adiponectin receptor protein
UniProt Gene Name:Adipor1  
UniProt Entry Name:ADR1_MOUSE
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Additional Information

Reactivity:
Mouse
ELISA Type:
Sandwich
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