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Human BMP-2 (Bone Morphogenetic Protein 2) ELISA Kit
Synonyms:BMP2A; BMP-2A; BDA2

Principle

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 Human BMP-2 . 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 Human BMP-2 and Avidin-Horseradish Peroxidase (HRP) conjugate is added to each micro plate well successively and then incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human BMP-2 , biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color turns to 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 Human BMP-2 . You can calculate the concentration of Human BMP-2 in the samples by comparing the OD of the samples to the standard curve.

Product ID E-EL-H0011
Assay type Sandwich ELISA
Format 96T
Assay time 4.5h
Reactivity Human
Detection Method Colormetric
Detection Range 62.500—4000.000pg/mL
Sensitivity 37.500pg/mL
Sample Volume Required Per Well 100μL
Sample Type serum, plasma and other biological fluids

Components

Item Specifications Storage
Micro ELISA Plate 8 wells ×12 strips 4℃/-20℃#
Reference Standard 2 vials 4℃/-20℃#
Reference Standard & Sample Diluent 1vial 20mL 4℃
Concentrated Biotinylated Detection Ab 1vial 120μL 4℃/-20℃#
Biotinylated Detection Ab Diluent 1vial 10mL 4℃
Concentrated HRP Conjugate 1vial 120μL 4℃(shading light)
HRP Conjugate Diluent 1vial 10mL 4℃
Concentrated Wash Buffer (25×) 1vial 30mL 4℃
Substrate Reagent 1vial 10mL 4℃(shading light)
 Stop Solution 1vial 10mL 4℃
Plate Sealer 5pieces
Manual 1 copy
Certificate of Analysis 1 copy
#: keep the kit at 4℃ if it’s used within 30 days, keep at -20℃ for longer storage.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Human BMP-2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human BMP-2 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
(pg/mL)
199.900 617.200 1476.000 215.600 594.500 1427.300
Standard deviation 10.400 25.900 45.800 11.000 25.000 47.100
C V (%) 5.200 4.200 3.100 5.100 4.200 3.300

Recovery

The recovery of Human BMP-2 spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.

Sample Type Range (%) Average Recovery (%)
Serum (n=5) 87-97 92.000
EDTA plasma (n=5) 84-97 91.000
Cell culture media (n=5) 85-95 90.000

Linearity

Samples were spiked with high concentrations of Human BMP-2 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)
1:2 Range (%) 87-100 86-102 97-108
Average (%) 92.000 93.000 103.000
1:4 Range (%) 97-112 80-94 91-104
Average (%) 105.000 87.000 98.000
1:8 Range (%) 98-114 81-94 91-108
Average (%) 106.000 86.000 98.000
1:16 Range (%) 94-110 83-94 96-106
Average (%) 100.000 89.000 101.000

Specificity

This kit recognizes recombinant and natural Human BMP-2 . No significant cross-reactivity or interference was observed.

Procedure

  1. Add 100μL standard or sample to each well. Incubate 90 mintues at 37℃.
  2. Add 100μL Biotinylated Detection Ab. Incubate 1 hour at 37℃.
  3. Aspirate and wash 3 times.
  4. Add 100μL HRP Conjugate. Incubate 30 minutes at 37℃.
  5. Aspirate and wash 5 times.
  6. Add 90μL Substrate Reagent. Incubate 15 minutes at 37℃.
  7. Add 50μL Stop Solution. Read it at 450nm immediately.
  8. Calculation of results.

Storage

All the reagents in the kit should be stored according to the labels on vials. Unused wells should be returned to the foil pouch with the desiccant pack and resealed along entire edge of zip-seal. Substrate Reagent shouldn't be kept at -20℃ (Check!). Exposure of reagents to strong light should be avoided in the process of incubation and storage. All the taps of reagents should be tightened to prevent evaporation and microbial contamination.

If you need the manual for more details, you can click here! 

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