Mouse Slc18a2(Solute Carrier Family 18,Member 2)ELISA Kit (MOES01759)

SKU:
MOES01759
€549

Description

ELISA Kit Technical ManualMSDS

Mouse Slc18a2(Solute Carrier Family 18,Member 2)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 Slc18a2 . 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 Slc18a2 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 Slc18a2, 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 Slc18a2. The concentration of Mouse Slc18a2 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 Range78.13-5000 pg/mL
Sensitivity46.88 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Mouse Slc18a2 in samples. No significant cross-reactivity or interference between Mouse Slc18a2 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
50002.447
2.463
2.4552.397
25001.731
1.733
1.7321.674
12501.034
1.004
1.0190.961
6250.466
0.5
0.4830.425
312.50.284
0.258
0.2710.213
156.250.173
0.163
0.1680.11
78.130.111
0.117
0.1140.056
00.052
0.064
0.058--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse Slc18a2 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse Slc18a2 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)270.84438.062421.68259.43413.282370.96
Standard deviation14.0618.22114.7917.8718.10105.98
C V (%)5.194.164.746.894.384.47

Recovery

The recovery of Mouse Slc18a2 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)94-107101
EDTA plasma (n=5)91-10597
Cell culture media (n=5)89-10396

Linearity

Samples were spiked with high concentrations of Mouse Slc18a2 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 (%)96-10990-10090-102
Average (%)1039596
1:4Range (%)93-10880-9385-97
Average (%)998691
1:8Range (%)93-10885-9786-100
Average (%)999093
1:16Range (%)91-10885-10089-102
Average (%)989294

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 Slc18a2(Solute Carrier Family 18,Member 2)ELISA Kit (MOES01759) 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 Slc18a2(Solute Carrier Family 18,Member 2)ELISA Kit (MOES01759) Protein Information

UniProt Protein Function:SLC18A2: Involved in the ATP-dependent vesicular transport of biogenic amine neurotransmitters. Pumps cytosolic monoamines including dopamine, norepinephrine, serotonin, and histamine into synaptic vesicles. Requisite for vesicular amine storage prior to secretion via exocytosis. Belongs to the major facilitator superfamily. Vesicular transporter family.
UniProt Protein Details:

Protein type:Transporter; Transporter, SLC family; Membrane protein, multi-pass; Membrane protein, integral

Chromosomal Location of Human Ortholog: 10q25

Cellular Component: synaptic vesicle; synaptic vesicle membrane; membrane; cell soma; integral to plasma membrane; plasma membrane; terminal button

Molecular Function:serotonin transmembrane transporter activity; enzyme binding; amine transmembrane transporter activity; heat shock protein binding; monoamine transmembrane transporter activity; drug binding

Biological Process: dopamine transport; neurotransmitter secretion; death; amine transport; response to amphetamine; monoamine transport; locomotory behavior; response to herbicide; glucose homeostasis; endocytic recycling; response to cocaine; sequestering of neurotransmitter; post-embryonic development; response to starvation; synaptic transmission; insulin secretion; response to zinc ion; serotonin transport; transmembrane transport; negative regulation of neurotransmitter transport; aging; response to corticosterone stimulus

NCBI Summary:The vesicular monoamine transporter acts to accumulate cytosolic monoamines into synaptic vesicles, using the proton gradient maintained across the synaptic vesicular membrane. Its proper function is essential to the correct activity of the monoaminergic systems that have been implicated in several human neuropsychiatric disorders. The transporter is a site of action of important drugs, including reserpine and tetrabenazine (summary by Peter et al., 1993 [PubMed 7905859]). See also SLC18A1 (MIM 193002).[supplied by OMIM, Jan 2011]
UniProt Code:Q8BRU6
NCBI GenInfo Identifier:1722742
NCBI Gene ID:6571
NCBI Accession:Q05940.2
UniProt Secondary Accession:Q8BRU6,Q8BRU6, Q01827,
UniProt Related Accession:Q05940
Molecular Weight:514
NCBI Full Name:Synaptic vesicular amine transporter
NCBI Synonym Full Names:solute carrier family 18 (vesicular monoamine transporter), member 2
NCBI Official Symbol:SLC18A2
NCBI Official Synonym Symbols:SVAT; SVMT; VAT2; VMAT2
NCBI Protein Information:synaptic vesicular amine transporter; monoamine transporter; vesicular amine transporter 2; vesicle monoamine/H+ antiporter; solute carrier family 18 member 2; vesicle monoamine transporter type 2; monoamine neurotransmitter transporter; solute carrier fa
UniProt Protein Name:Synaptic vesicular amine transporter
UniProt Synonym Protein Names:Monoamine transporter; Solute carrier family 18 member 2; Vesicular amine transporter 2; VAT2
UniProt Gene Name:SLC18A2
UniProt Entry Name:VMAT2_HUMAN
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Additional Information

Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
Sandwich
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