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Bovine Collagen Type III (Col III) ELISA Kit

Citations(0) Uniprot : N/A
  • Cat.No.: AE64731BO

  • Reactivity: Bovine (Bos taurus; Cattle)

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48T 96T
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Assay Procedure Summary







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Product Details

种属 Bovine (Bos taurus; Cattle)
UniProt号 N/A
缩写 Col III
别名 N/A
线性范围 6.25-400 ng/mL
灵敏度 3.12 ng/mL
样本类型 Serum, Plasma, Other biological fluids
检测方法 Sandwich
分析方法 Quantitive
反应时间 1-4.5h
样本体积 1-200 μL
检测波长 450 nm

原理

This assay employs a two-site sandwich ELISA to quantitate Col III in samples. An antibody specific for Col III has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Col III present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for Col III 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 Col III bound in the initial step. The color development is stopped and the intensity of the color is measured.
 

产品概述


 

组份


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

 

特异性

This assay has high sensitivity and excellent specificity for detection of Bovine Col III. No significant cross-reactivity or interference between Bovine Col III and analogues was observed.
 

回收率

Matrices listed below were spiked with certain level of recombinant Bovine Col III and the recovery rates were calculated by comparing the measured value to the expected amount of Bovine Col III 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

 

精密度

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%
 

线性

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Bovine Col III 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%

 

稳定性

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).
 

样本收集及储存

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.
 

注意事项

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