Mouse BDNF (Brain Derived Neurotrophic Factor) ELISA Kit (MOES00782)

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MOES00782
€299

Description

ELISA Kit Technical ManualMSDS

Mouse BDNF (Brain Derived Neurotrophic Factor) ELISA Kit

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 Mouse BDNF . 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 Mouse BDNF 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 BDNF, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns 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 Mouse BDNF. The concentration of Mouse BDNF in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
Detection MethodColormetric
Detection Range31.25-2000 pg/mL
Sensitivity18.75 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

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

Typical Data

As the OD 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)
O.D Average Corrected
20002.422
2.468
2.4452.355
10001.561
1.605
1.5831.493
5001.003
0.965
0.9840.894
2500.486
0.49
0.4880.398
1250.281
0.265
0.2730.183
62.50.194
0.188
0.1910.101
31.250.138
0.146
0.1420.052
00.082
0.098
0.09--

Precision

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

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

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)108.30200.70906.70117.20182.00864.00
Standard deviation7.509.0029.006.607.6035.40
C V (%)6.934.483.205.634.184.10

Recovery

The recovery of Mouse BDNF 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)93-10799
EDTA plasma (n=5)93-109100
Cell culture media (n=5)95-111101

Linearity

Samples were spiked with high concentrations of Mouse BDNF 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 (%)87-9886-9694-109
Average (%)9291101
1:4Range (%)98-11080-9292-107
Average (%)10386100
1:8Range (%)97-11084-9797-111
Average (%)10591103
1:16Range (%)99-11482-9795-109
Average (%)10788102

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 ELISA 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 Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Mouse BDNF (Brain Derived Neurotrophic Factor) ELISA Kit (MOES00782) 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 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Mouse BDNF (Brain Derived Neurotrophic Factor) ELISA Kit (MOES00782) Protein Information

UniProt Protein Function:BDNF: During development, promotes the survival and differentiation of selected neuronal populations of the peripheral and central nervous systems. Participates in axonal growth, pathfinding and in the modulation of dendritic growth and morphology. Major regulator of synaptic transmission and plasticity at adult synapses in many regions of the CNS. The versatility of BDNF is emphasized by its contribution to a range of adaptive neuronal responses including long-term potentiation (LTP), long-term depression (LTD), certain forms of short-term synaptic plasticity, as well as homeostatic regulation of intrinsic neuronal excitability. Defects in BDNF are a cause of congenital central hypoventilation syndrome (CCHS); also known as congenital failure of autonomic control or Ondine curse. CCHS is a rare disorder characterized by abnormal control of respiration in the absence of neuromuscular or lung disease, or an identifiable brain stem lesion. A deficiency in autonomic control of respiration results in inadequate or negligible ventilatory and arousal responses to hypercapnia and hypoxemia. CCHS is frequently complicated with neurocristopathies such as Hirschsprung disease that occurs in about 16% of CCHS cases. Belongs to the NGF-beta family. 5 isoforms of the human protein are produced by alternative promoter.
UniProt Protein Details:

Protein type:Secreted; Cytokine; Cell development/differentiation; Secreted, signal peptide

Cellular Component: extracellular space; cell; cytoplasmic membrane-bound vesicle; dendrite; extracellular region; terminal button; perikaryon; synaptic vesicle; mitochondrial crista; cell soma; perinuclear region of cytoplasm; axon; cytoplasm

Molecular Function:protein binding; growth factor activity; neurotrophin TRKB receptor binding; receptor binding

Biological Process: regulation of long-term neuronal synaptic plasticity; fear response; axon guidance; mechanoreceptor differentiation; behavioral fear response; mitochondrial electron transport, NADH to ubiquinone; axon extension; regulation of synaptic plasticity; positive regulation of long-term neuronal synaptic plasticity; regulation of inhibitory postsynaptic membrane potential; learning and/or memory; negative regulation of neuroblast proliferation; ureteric bud development; regulation of neuron apoptosis; dendrite development; regulation of retinal cell programmed cell death; feeding behavior; regulation of metabolic process; negative regulation of neuron apoptosis; negative regulation of synaptic transmission, GABAergic; inner ear development; response to drug; nervous system development; neuron recognition; regulation of axon extension; regulation of short-term neuronal synaptic plasticity; learning; negative regulation of striated muscle development; positive regulation of peptidyl-serine phosphorylation; positive regulation of synaptogenesis; axon target recognition; glutamate secretion; positive regulation of transcription factor activity; nerve development; regulation of excitatory postsynaptic membrane potential; positive regulation of neuron differentiation; gamma-aminobutyric acid signaling pathway; transmembrane receptor protein tyrosine kinase signaling pathway; negative regulation of apoptosis

NCBI Summary:The protein encoded by this gene is a member of the nerve growth factor family. It is involved in the growth, differentiation and survival of specific types of developing neurons both in the central nervous system (CNS) and the peripheral nervous system. It is also involved in regulating synaptic plasticity in the CNS. Expression of a similar gene in human is reduced in both Alzheimer's and Huntington disease patients. Multiple transcript variants encoding distinct isoforms have been described for this gene. [provided by RefSeq, Sep 2015]
UniProt Code:P21237
NCBI GenInfo Identifier:114901
NCBI Gene ID:12064
NCBI Accession:P21237.1
UniProt Related Accession:P21237
Molecular Weight:28,123 Da
NCBI Full Name:Brain-derived neurotrophic factor
NCBI Synonym Full Names:brain derived neurotrophic factor
NCBI Official Symbol:Bdnf  
NCBI Protein Information:brain-derived neurotrophic factor; anorexia BDNF
UniProt Protein Name:Brain-derived neurotrophic factor
Protein Family:BDNF/NT-3 growth factors receptor
UniProt Gene Name:Bdnf  
UniProt Entry Name:BDNF_MOUSE
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Additional Information

Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
Sandwich
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