Mouse Cell Signalling ELISA Kits 2
Mouse BDNF (Brain Derived Neurotrophic Factor) CLIA Kit (MOES00129)
- SKU:
- MOES00129
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 7.5pg/mL
- Range:
- 12.5-800pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- ANON2, BULN2
- Reactivity:
- Mouse
- Sample Type:
- Serum, plasma and other biological fluids
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection method: | Chemiluminescence |
| Detection range: | 12.50-800 pg/mL |
| Sensitivity: | 7.50 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Mouse BDNF in samples. No significant cross-reactivity or interference between Mouse BDNF and analogues was observed. |
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 BDNF. 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 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 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 BDNF. The concentration of Mouse BDNF in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| 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 |
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 |
| 800 | 47876 57408 | 52642 | 52616 |
| 400 | 20440 23028 | 21734 | 21708 |
| 200 | 10018 9398 | 9708 | 9682 |
| 100 | 4441 4663 | 4552 | 4526 |
| 50 | 2321 2055 | 2188 | 2162 |
| 25 | 1129 991 | 1060 | 1034 |
| 12.50 | 498 520 | 509 | 483 |
| 0 | 26 26 | 26 | -- |
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 | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 42.80 | 106.52 | 276.31 | 44.72 | 111.91 | 267.35 |
| Standard deviation | 3.94 | 12.27 | 18.43 | 5.65 | 11.47 | 28.42 |
| C V (%) | 9.21 | 11.52 | 6.67 | 12.63 | 10.25 | 10.63 |
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) | 88-100 | 94 |
| EDTA plasma (n=5) | 99-115 | 107 |
| Cell culture media (n=5) | 95-105 | 100 |
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) | ||
| 1:2 | Range (%) | 95-111 | 87-103 | 97-112 |
| Average (%) | 103 | 94 | 103 | |
| 1:4 | Range (%) | 90-102 | 94-110 | 94-109 |
| Average (%) | 96 | 101 | 101 | |
| 1:8 | Range (%) | 96-109 | 95-110 | 86-100 |
| Average (%) | 103 | 100 | 93 | |
| 1:16 | Range (%) | 100-115 | 96-111 | 94-108 |
| Average (%) | 108 | 103 | 101 |
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 ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- 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.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- 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.
- 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.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- 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.
- Determine the RLU value of each well immediately.
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