Human SNC alpha (Synuclein Alpha) 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 Human SNC alpha . 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 SNC alpha 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 Human SNC alpha, 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 Human SNC alpha. The concentration of Human SNC alpha in samples can be calculated by comparing the OD of the samples to the standard curve.
|Detection Range||15.63-1000 pg/mL|
|Sample Volume Required Per Well||100uL|
|Sample Type||Serum, plasma and other biological fluids|
This kit recognizes Human SNC alpha in samples. No significant cross-reactivity or interference between Human SNC alpha and analogues was observed.
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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human SNC alpha were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human SNC alpha were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||6.79||5.81||3.10||6.57||5.19||4.84|
The recovery of Human SNC alpha spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||92-108||99|
|Cell culture media (n=5)||90-101||96|
Samples were spiked with high concentrations of Human SNC alpha 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)|
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.
|Micro ELISA Plate(Dismountable)||8 wells X12 strips||-20'C, 6 months|
|Reference Standard||2 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 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 (25X)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4'C(shading light)|
|Stop Solution||1 vial, 10 mL||4'C|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|
Human SNC alpha (Synuclein Alpha) ELISA Kit (HUES02100) 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 100 µL of standard solutions into the standard wells.
- 3. Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- 4. Add 100 µL 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 100 µL 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 350 µL 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 100 µL 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 90 µL 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.
Human SNC alpha (Synuclein Alpha) ELISA Kit (HUES02100) Protein Information
|UniProt Protein Function:||SNCA: a member of the synuclein family. Abundantly expressed in the brain. Inhibits phospholipase D2 selectively. May integrate presynaptic signaling and membrane trafficking. Implicated in the pathogenesis of Parkinson's disease. A major component of amyloid plaques in the brains of patients with Alzheimer's disease. Two alternatively spliced isoforms transcripts have been identified.|
|UniProt Protein Details:|
Chromosomal Location of Human Ortholog: 4q21
Cellular Component: Golgi apparatus; nuclear outer membrane; mitochondrion; rough endoplasmic reticulum; lysosome; extracellular region; fibril; terminal button; inclusion body; cell cortex; mitochondrial respiratory chain complex I; cytosol; actin cytoskeleton; synaptic vesicle; platelet alpha granule membrane; growth cone; perinuclear region of cytoplasm; axon; cytoplasm; plasma membrane; ribosome; cell junction; nucleus
Molecular Function:protein domain specific binding; identical protein binding; histone binding; zinc ion binding; kinesin binding; ferrous iron binding; microtubule binding; caspase inhibitor activity; beta-tubulin binding; magnesium ion binding; phosphoprotein binding; protein N-terminus binding; oxidoreductase activity; Hsp70 protein binding; calcium ion binding; dynein binding; protein binding; copper ion binding; phospholipase binding; phospholipid binding; tau protein binding; fatty acid binding; alpha-tubulin binding
Biological Process: regulation of long-term neuronal synaptic plasticity; negative regulation of serotonin uptake; regulation of acyl-CoA biosynthetic process; adult locomotory behavior; positive regulation of apoptosis; negative regulation of norepinephrine uptake; mitochondrial membrane organization and biogenesis; microglial cell activation; response to lipopolysaccharide; positive regulation of endocytosis; dopamine biosynthetic process; negative regulation of transcription from RNA polymerase II promoter; negative regulation of caspase activity; negative regulation of monooxygenase activity; fatty acid metabolic process; positive regulation of neurotransmitter secretion; regulation of dopamine secretion; negative regulation of dopamine uptake; negative regulation of histone acetylation; calcium ion homeostasis; negative regulation of exocytosis; response to magnesium ion; negative regulation of protein amino acid phosphorylation; behavioral response to cocaine; receptor internalization; phospholipid metabolic process; fibril organization and biogenesis; synapse organization and biogenesis; dopamine uptake; negative regulation of neuron apoptosis; response to iron(II) ion; positive regulation of receptor recycling; aging; caspase activation; response to drug; neutral lipid metabolic process; protein destabilization; regulation of macrophage activation; regulation of glutamate secretion; negative regulation of microtubule polymerization; positive regulation of peptidyl-serine phosphorylation; negative regulation of dopamine metabolic process; organelle ATP synthesis coupled electron transport; regulation of locomotion; synaptic vesicle endocytosis; positive regulation of release of sequestered calcium ion into cytosol; regulation of excitatory postsynaptic membrane potential; negative regulation of transporter activity; negative regulation of apoptosis
Disease: Parkinson Disease 4, Autosomal Dominant; Dementia, Lewy Body; Parkinson Disease 1, Autosomal Dominant
|NCBI Summary:||Alpha-synuclein is a member of the synuclein family, which also includes beta- and gamma-synuclein. Synucleins are abundantly expressed in the brain and alpha- and beta-synuclein inhibit phospholipase D2 selectively. SNCA may serve to integrate presynaptic signaling and membrane trafficking. Defects in SNCA have been implicated in the pathogenesis of Parkinson disease. SNCA peptides are a major component of amyloid plaques in the brains of patients with Alzheimer's disease. Four alternatively spliced transcripts encoding two different isoforms have been identified for this gene. [provided by RefSeq, Mar 2009]|
|NCBI GenInfo Identifier:||586067|
|NCBI Gene ID:||6622|
|UniProt Secondary Accession:||P37840,Q13701, Q4JHI3, Q6IAU6, A8K2A4,|
|UniProt Related Accession:||P37840|
|NCBI Full Name:||Alpha-synuclein|
|NCBI Synonym Full Names:||synuclein, alpha (non A4 component of amyloid precursor)|
|NCBI Official Symbol:||SNCA|
|NCBI Official Synonym Symbols:||PD1; NACP; PARK1; PARK4|
|NCBI Protein Information:||alpha-synuclein; synuclein alpha-140; non A-beta component of AD amyloid|
|UniProt Protein Name:||Alpha-synuclein|
|UniProt Synonym Protein Names:||Non-A beta component of AD amyloid; Non-A4 component of amyloid precursor; NACP|
|UniProt Gene Name:||SNCA|
|UniProt Entry Name:||SYUA_HUMAN|