Mouse BMP-1 (Bone Morphogenetic Protein 1) CLIA Kit (MOES00121)

SKU:
MOES00121
Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
CLIA
€649

Description

ELISA Kit Technical ManualMSDS

Mouse BMP-1 (Bone Morphogenetic Protein 1) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Mouse BMP-1 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse BMP-1 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 BMP-1, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Mouse BMP-1. The concentration of Mouse BMP-1 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
Detection methodChemiluminescence
Detection range15.63-1000 pg/mL
Sensitivity9.38 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Mouse BMP-1 in samples. No significant cross-reactivity or interference between Mouse BMP-1 and analogues was observed.

Typical Data

As the RLU 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
(pg/mL)
RLU Average Corrected
100050105
57245
5367553650
50020788
24190
2248922464
25011015
9407
1021110186
1254667
5135
49014876
62.52547
2359
24532428
31.251297
1265
12811256
15.63660
756
708683
024
26
25--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse BMP-1 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse BMP-1 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)49.69124.12414.0051.01120.12394.80
Standard deviation4.3910.8131.055.9113.3637.19
C V (%)8.838.717.5011.5911.129.42

Recovery

The recovery of Mouse BMP-1 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)93-109100
EDTA plasma (n=5)86-9991
Cell culture media (n=5)101-115106

Linearity

Samples were spiked with high concentrations of Mouse BMP-1 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 (%)86-9793-10596-110
Average (%)9298102
1:4Range (%)85-10088-10295-109
Average (%)9295102
1:8Range (%)87-10087-10096-109
Average (%)9392103
1:16Range (%)90-10384-9990-105
Average (%)969197

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 CLIA 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 Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Mouse BMP-1 (Bone Morphogenetic Protein 1) CLIA Kit (MOES00121) 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 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
    11. Determine the RLU value of each well immediately.

Mouse BMP-1 (Bone Morphogenetic Protein 1) CLIA Kit (MOES00121) Protein Information

UniProt Protein Function:BMP1: Cleaves the C-terminal propeptides of procollagen I, II and III. Induces cartilage and bone formation. May participate in dorsoventral patterning during early development by cleaving chordin (CHRD). Defects in BMP1 are a cause of autosomal recessive osteogenesis imperfecta (AR-OI). A connective tissue disorder characterized by bone fragility, progressively deforming bones, bowing of limbs due to multiple fractures, very short stature, a triangular face, severe scoliosis, and grayish sclera. AR-OI due to BMP1 mutations belongs to the group of osteogenesis imperfecta type III in the Sillence classification. Belongs to the peptidase M12A family. 7 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Protease; Cytokine; EC 3.4.24.19

Cellular Component: Golgi apparatus; extracellular space; proteinaceous extracellular matrix; extracellular region

Molecular Function:peptidase activity; protein binding; growth factor activity; zinc ion binding; hydrolase activity; metallopeptidase activity; metalloendopeptidase activity; metal ion binding; cytokine activity; calcium ion binding

Biological Process: ossification; cartilage development; multicellular organismal development; proteolysis; cell differentiation

NCBI Summary:This gene encodes a metalloproteinase that plays an essential role in the formation of the extracellular matrix and is also able to induce ectopic bone formation. Unlike other bone morphogenetic proteins, the protein encoded by this gene is not closely related to transforming growth factor-beta. This protein plays in role several developmental processes. In humans, mutations in this gene are associated with osteogenesis imperfecta and with increased bone mineral density and multiple recurrent fractures. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Dec 2013]
UniProt Code:P98063
NCBI GenInfo Identifier:42734447
NCBI Gene ID:12153
NCBI Accession:NP_033885.2
UniProt Secondary Accession:P98063,Q6NZM2,
UniProt Related Accession:P98063
Molecular Weight:111,666 Da
NCBI Full Name:bone morphogenetic protein 1
NCBI Synonym Full Names:bone morphogenetic protein 1
NCBI Official Symbol:Bmp1  
NCBI Official Synonym Symbols:Pcp; Tld  
NCBI Protein Information:bone morphogenetic protein 1
UniProt Protein Name:Bone morphogenetic protein 1
UniProt Synonym Protein Names:Mammalian tolloid protein; mTld; Procollagen C-proteinase; PCP
Protein Family:Bone morphogenetic protein
UniProt Gene Name:Bmp1  
UniProt Entry Name:BMP1_MOUSE
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