Cat NADH-ubiquinone oxidoreductase chain 1 (MT-ND1) ELISA Kit

Cat# AE32409CA-48

Size : 48Tests

Brand : Abebio

Contact local distributor :


Phone : +1 850 650 7790

Reagent Preparation







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Assay Procedure Summary







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Product Details

Species Reactivity Cat (Felis catus,Feline)
UniProt P48900
Abbreviation MT-ND1
Alternative Names MTND1; NADH dehydrogenase subunit 1
Range 62.5-4000 pg/mL
Sensitivity 31.25 pg/mL
Sample Type Serum, Plasma, Other biological fluids
Detection Method Sandwich
Analysis Method Quantitive
Assay Duration 1-4.5h
Sample Volume 1-200 μL
Detection Wavelengt 450 nm

Test principle

This assay employs a two-site sandwich ELISA to quantitate MT-ND1 in samples. An antibody specific for MT-ND1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MT-ND1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for MT-ND1 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of MT-ND1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
 

Product Overview


 

Components


Reagents

Quantity

Reagents

Quantity

Assay plate (96 Wells)

1

Instruction manual

1

Standard (lyophilized)

2

Sample Diluent

1 x 20 mL

Biotin-Conjugate (concentrate 100 x)

1 x 120 μL

Biotin-Conjugate Diluent

1 x 20 mL

Streptavidin-HRP  (concentrate 100 x)

1 x 120 μL

Streptavidin-HRP Diluent   

1 x 20 mL

Wash Buffer (concentrate 25 x)

1 x 20 mL

Substrate Solution

1 x 10 mL

Stop Solution

1 x 10 mL

Adhesive Films

4

 

Specificity

This assay has high sensitivity and excellent specificity for detection of Cat MT-ND1. No significant cross-reactivity or interference between Cat MT-ND1 and analogues was observed.
 

Recovery

Matrices listed below were spiked with certain level of recombinant Cat MT-ND1 and the recovery rates were calculated by comparing the measured value to the expected amount of Cat MT-ND1 in samples.

Sample Type

Number

Recovery range (%)

Average(%)

Serum

10

90-101

96

EDTA plasma

10

89-97

93

Heparin plasma

10

91-99

95

 

Precision

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
 

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Cat MT-ND1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample Type

1:2

1:4

1:8

1:16

Serum

78-89%

81-99%

92-103%

95-105%

EDTA plasma

91-101%

90-98%

93-101%

91-98%

Heparin plasma

92-103%

93-102%

92-99%

91-101%

 

Stability

The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
 

Sample collection and storage

Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
 

Kits storage instructions

Store at 2-8°C. Please refer to Instruction Manual.