Human PS2 (Presenilin 2) ELISA Kit (HUES03156)

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HUES03156
€539

Description

ELISA Kit Technical ManualMSDS

Human PS2 (Presenilin 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 Human PS2 . 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 PS2 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 PS2, 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 PS2. The concentration of Human PS2 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
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 Human PS2 in samples. No significant cross-reactivity or interference between Human PS2 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
20002.488
2.528
2.5082.447
10001.561
1.565
1.5631.502
5000.93
0.896
0.9130.852
2500.47
0.474
0.4720.411
1250.244
0.228
0.2360.175
62.50.173
0.151
0.1620.101
31.250.106
0.12
0.1130.052
00.055
0.067
0.061--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PS2 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PS2 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)110.22166.70973.77116.84174.871030.00
Standard deviation6.849.8936.427.588.4855.00
C V (%)6.215.933.746.494.855.34

Recovery

The recovery of Human PS2 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)86-9691
EDTA plasma (n=5)92-10698
Cell culture media (n=5)94-109100

Linearity

Samples were spiked with high concentrations of Human PS2 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 (%)93-11194-10698-115
Average (%)10199105
1:4Range (%)92-10784-9881-95
Average (%)989088
1:8Range (%)87-9886-9981-95
Average (%)939387
1:16Range (%)92-10786-9987-100
Average (%)989193

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

Human PS2 (Presenilin 2) ELISA Kit (HUES03156) 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.

Human PS2 (Presenilin 2) ELISA Kit (HUES03156) Protein Information

UniProt Protein Function:PSEN2: probable catalytic subunit of the gamma-secretase complex, an endoprotease complex that catalyzes the intramembrane cleavage of integral membrane proteins such as Notch receptors and APP (beta-amyloid precursor protein). An integral membrane protein of the Golgi and endoplasmic reticulum. Requires the other members of the gamma-secretase complex to have a protease activity. May play a role in intracellular signaling and gene expression or in linking chromatin to the nuclear membrane. May function in the cytoplasmic partitioning of proteins. Two alternative splice isoforms have been identified.
UniProt Protein Details:

Protein type:EC 3.4.23.-; Membrane protein, integral; Membrane protein, multi-pass; Cell surface; Protease

Chromosomal Location of Human Ortholog: 1q42.13

Cellular Component: Golgi apparatus; centrosome; cell surface; integral to plasma membrane; lysosomal membrane; nuclear inner membrane; cytosol; lipid raft; ciliary rootlet; kinetochore; cell soma; membrane; perinuclear region of cytoplasm; apical plasma membrane; mitochondrial inner membrane; dendritic shaft; neuromuscular junction; endoplasmic reticulum membrane; protein complex; endoplasmic reticulum; cell cortex; Z disc; Golgi membrane; growth cone; axon; plasma membrane

Molecular Function:protein binding; endopeptidase activity; aspartic-type endopeptidase activity

Biological Process: positive regulation of catalytic activity; axon guidance; nerve growth factor receptor signaling pathway; positive regulation of apoptosis; protein maturation; regulation of synaptic plasticity; positive regulation of coagulation; cell fate specification; T cell receptor signaling pathway; T cell activation during immune response; protein transport; amyloid precursor protein catabolic process; hair follicle development; negative regulation of protein amino acid phosphorylation; beta-amyloid metabolic process; calcium ion transport; forebrain development; ephrin receptor signaling pathway; brain morphogenesis; negative regulation of protein binding; dorsoventral neural tube patterning; embryonic limb morphogenesis; cardiac muscle contraction; Notch signaling pathway; somitogenesis; membrane protein intracellular domain proteolysis; thymus development; regulation of epidermal growth factor receptor activity; membrane protein ectodomain proteolysis; Notch receptor processing; memory; endoplasmic reticulum calcium ion homeostasis; myeloid leukocyte differentiation; response to hypoxia; hemopoietic progenitor cell differentiation; protein processing; negative regulation of protein complex assembly; alveolus development; negative regulation of apoptosis

Disease: Alzheimer Disease 4; Cardiomyopathy, Dilated, 1v

NCBI Summary:Alzheimer's disease (AD) patients with an inherited form of the disease carry mutations in the presenilin proteins (PSEN1 or PSEN2) or the amyloid precursor protein (APP). These disease-linked mutations result in increased production of the longer form of amyloid-beta (main component of amyloid deposits found in AD brains). Presenilins are postulated to regulate APP processing through their effects on gamma-secretase, an enzyme that cleaves APP. Also, it is thought that the presenilins are involved in the cleavage of the Notch receptor such that, they either directly regulate gamma-secretase activity, or themselves act are protease enzymes. Two alternatively spliced transcript variants encoding different isoforms of PSEN2 have been identified. [provided by RefSeq, Jul 2008]
UniProt Code:P49810
NCBI GenInfo Identifier:1709858
NCBI Gene ID:5664
NCBI Accession:P49810.1
UniProt Secondary Accession:P49810,Q96P32, A8K8D4, B1AP21,
UniProt Related Accession:P49810
Molecular Weight:50,011 Da
NCBI Full Name:Presenilin-2
NCBI Synonym Full Names:presenilin 2
NCBI Official Symbol:PSEN2  
NCBI Official Synonym Symbols:AD4; PS2; AD3L; STM2; CMD1V  
NCBI Protein Information:presenilin-2; AD5; E5-1; PS-2; AD3LP; STM-2; Alzheimer disease 4
UniProt Protein Name:Presenilin-2
UniProt Synonym Protein Names:AD3LP; AD5; E5-1; STM-2Cleaved into the following 2 chains:Presenilin-2 NTF subunit; Presenilin-2 CTF subunit
Protein Family:Presenilin
UniProt Gene Name:PSEN2  
UniProt Entry Name:PSN2_HUMAN
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Additional Information

Reactivity:
Human
ELISA Type:
Sandwich
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