Pig Mycoplasma pneumoniae antibody (MP-Ab) ELISA Kit

Referência AE62681PI

Tamanho : 96Tests

Marca : Abebio

Solicitar mais informações

Contactar o distribuidor local :


Telefone : +1 850 650 7790

Reagent Preparation







Results demonstration







Assay Procedure Summary







Certificate



Product Details

Species Reactivity Pig (Sus scrofa; Porcine)
UniProt
Abbreviation MP-Ab
Alternative Names N/A
Range Request Information
Sensitivity Request Information
Sample Type Serum, Plasma, Other biological fluids
Detection Method Competitive ELISA
Analysis Method Qualitative
Assay Duration 1-3h
Sample Volume 1-200 μL
Detection Wavelengt 450 nm

Test principle

This assay employs the competitive enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to MP-Ab. Standards or samples are then added to the appropriate microtiter plate wells with a Horseradish Peroxidase (HRP)-conjugated MP-Ab and incubated. The competitive inhibition reaction is launched between with HRP labeled MP-Ab and unlabeled MP-Ab with the antibody. A substrate solution is added to the wells and the color develops in opposite to the amount of MP-Ab in the sample. 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 12 mL

Streptavidin-HRP  (concentrate 100 x)

1 x 120 μL

Streptavidin-HRP Diluent   

1 x 12 mL

Wash Buffer (concentrate 25 x)

1 x 20 mL

Substrate Solution

1 x 10 mL

Stop Solution

1 x 6 mL

Adhesive Films

4

 

Specificity

This assay has high sensitivity and excellent specificity for detection of Pig MP-Ab. No significant cross-reactivity or interference between Pig MP-Ab and analogues was observed.
 

Recovery

Matrices listed below were spiked with certain level of recombinant Pig MP-Ab and the recovery rates were calculated by comparing the measured value to the expected amount of Pig MP-Ab 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 Pig MP-Ab 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.