人間 Presenilin-1 ELISA Kit (HUFI07066)- 高感度

商品名:	Human Presenilin-1 ELISA Kit
製品コード:	HUFI07066
サイズ:	96T
エイリアス:	AD3 ELISA Kit, FAD ELISA Kit, Presenilin-1 ELISA Kit, Protein S182 ELISA Kit, PS 1 ELISA Kit, PS1 ELISA Kit, PS1 CTF12 ELISA Kit, PSEN1 ELISA Kit, PSNL1 ELISA Kit, S182 ELISA Kit
検出方法:	Sandwich ELISA, Double Antibody
申し込み:	This immunoassay kit allows for the in vitro quantitative determination of Human Presenilin-1 concentrations in serum plasma and other biological fluids.
感度:	< 0.188ng/ml
範囲:	0.313-20ng/ml
保管所:	4°C for 6 months
ノート:	For Research Use Only

回復:	Matrices listed below were spiked with certain level of Human Presenilin-1 and the recovery rates were calculated by comparing the measured value to the expected amount of Human Presenilin-1 in samples. Enquire for more information.
直線性:	The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human Presenilin-1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Enquire for more information.
CV(%):	Intra-Assay: CV<8% Inter-Assay: CV<10%

成分	量	保管所
ELISA Microplate (Dismountable)	8×12 strips	4°C for 6 months
Lyophilized Standard	2	4°C/-20°C
Sample/Standard Dilution Buffer	20ml	4°C
Biotin-labeled Antibody(Concentrated)	120ul	4°C (Protect from light)
Antibody Dilution Buffer	10ml	4°C
HRP-Streptavidin Conjugate(SABC)	120ul	4°C (Protect from light)
SABC Dilution Buffer	10ml	4°C
TMB Substrate	10ml	4°C (Protect from light)
Stop Solution	10ml	4°C
Wash Buffer(25X)	30ml	4°C
Plate Sealer	5	-

必要なその他の材料と設備:

Microplate reader with 450 nm wavelength filter
Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
Incubator
Deionized or distilled water
Absorbent paper
Buffer resevoir

UniProt Protein Function:	PSEN1: 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). 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. Regulates epithelial- cadherin function. Five alternative splice isoforms have been identified.
UniProt Protein Details:	Protein type:Cell surface; Protease; Mitochondrial; Membrane protein, integral; Membrane protein, multi-pass; EC 3.4.23.- Chromosomal Location of Human Ortholog: 14q24.3 Cellular Component: Golgi apparatus; nuclear outer membrane; centrosome; cell surface; smooth endoplasmic reticulum; mitochondrion; lysosomal membrane; integral to plasma membrane; integral to membrane; lipid raft; ciliary rootlet; kinetochore; cell soma; membrane; perinuclear region of cytoplasm; mitochondrial inner membrane; apical plasma membrane; cytoplasmic vesicle; dendritic shaft; neuromuscular junction; endoplasmic reticulum membrane; nuclear membrane; rough endoplasmic reticulum; endoplasmic reticulum; cell cortex; Z disc; Golgi membrane; growth cone; axon Molecular Function:protein binding; cadherin binding; calcium channel activity; endopeptidase activity; beta-catenin binding; aspartic-type endopeptidase activity; PDZ domain binding Biological Process: positive regulation of catalytic activity; extracellular matrix organization and biogenesis; activation of MAPKK activity; positive regulation of apoptosis; beta-amyloid formation; regulation of synaptic plasticity; positive regulation of coagulation; Wnt receptor signaling pathway through beta-catenin; choline transport; T cell receptor signaling pathway; T cell activation during immune response; post-embryonic development; mitochondrial transport; extracellular matrix disassembly; positive regulation of MAP kinase activity; epithelial cell proliferation; cell-cell adhesion; negative regulation of axonogenesis; negative regulation of neuron apoptosis; embryonic limb morphogenesis; autophagic vacuole formation; somitogenesis; skin morphogenesis; regulation of phosphorylation; Notch receptor processing; negative regulation of ubiquitin-protein ligase activity; neuron development; Cajal-Retzius cell differentiation; hemopoietic progenitor cell differentiation; response to oxidative stress; negative regulation of apoptosis; regulation of synaptic transmission, glutamatergic; protein maturation; neuron migration; protein amino acid glycosylation; cell fate specification; myeloid dendritic cell differentiation; negative regulation of transcription from RNA polymerase II promoter; protein transport; amyloid precursor protein catabolic process; beta-amyloid metabolic process; regulation of resting membrane potential; brain morphogenesis; heart looping; positive regulation of receptor recycling; dorsoventral neural tube patterning; blood vessel development; Notch signaling pathway; L-glutamate transport; thymus development; membrane protein ectodomain proteolysis; memory; negative regulation of epidermal growth factor receptor activity; smooth endoplasmic reticulum calcium ion homeostasis; endoplasmic reticulum calcium ion homeostasis; neuron apoptosis; cerebral cortex cell migration; skeletal morphogenesis; regulation of protein binding; positive regulation of proteasomal ubiquitin-dependent protein catabolic process; protein processing; synaptic vesicle targeting; response to DNA damage stimulus Disease: Pick Disease Of Brain; Alzheimer Disease 3; Frontotemporal Dementia; Acne Inversa, Familial, 3; Cardiomyopathy, Dilated, 1u
NCBI Summary:	Alzheimer's disease (AD) patients with an inherited form of the disease carry mutations in the presenilin proteins (PSEN1; PSEN2) or in 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 are protease enzymes. Several alternatively spliced transcript variants encoding different isoforms have been identified for this gene, the full-length nature of only some have been determined. [provided by RefSeq, Aug 2008]
UniProt Code:	P49768
NCBI GenInfo Identifier:	1709856
NCBI Gene ID:	5663
NCBI Accession:	P49768.1
UniProt Secondary Accession:	P49768,O95465, Q14762, Q15719, Q15720, Q96P33, Q9UIF0 B2R6D3,
UniProt Related Accession:	P49768
Molecular Weight:	48,997 Da
NCBI Full Name:	Presenilin-1
NCBI Synonym Full Names:	presenilin 1
NCBI Official Symbol:	PSEN1
NCBI Official Synonym Symbols:	AD3; FAD; PS1; PS-1; S182
NCBI Protein Information:	presenilin-1
UniProt Protein Name:	Presenilin-1
UniProt Synonym Protein Names:	Protein S182Cleaved into the following 3 chains:Presenilin-1 NTF subunit; Presenilin-1 CTF subunit; Presenilin-1 CTF12; PS1-CTF12
Protein Family:	FHA domain-containing protein
UniProt Gene Name:	PSEN1
UniProt Entry Name:	PSN1_HUMAN

*ノート: プロトコルは、各バッチ/ロットに固有です。正しい手順については、キットに含まれているプロトコルに従ってください。

ウェルに加える前に、SABCワーキング溶液とTMB基質を37°Cで少なくとも30分間平衡化します。サンプルと試薬を希釈するときは、完全に均一に混合する必要があります。各テストの標準曲線をプロットすることをお勧めします。

ステップ	プロトコル
1.	Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.	Aliquot 0.1ml standard solutions into the standard wells.
3.	Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.	Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.	Seal the plate with a cover and incubate at 37 °C for 90 min.
6.	Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.	Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.	Seal the plate with a cover and incubate at 37 °C for 60 min.
9.	Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.	Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37 °C for 30 min.
11.	Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.	Add 90 µL of TMB substrate into each well, cover the plate and incubate at 37 °C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.	Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14.	Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

ELISAアッセイを実施する場合、可能な限り最良の結果を達成するためにサンプルを準備することが重要です。以下に、さまざまなサンプルタイプのサンプルを準備するための手順のリストを示します。

サンプルタイプ	プロトコル
血清	If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
プラズマ	Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
尿および脳脊髄液	Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
細胞培養上清	Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
細胞溶解物	Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
組織ホモジネート	The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
組織溶解物	Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
母乳	Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

Antibodies	ELISA Kits
Anti-Presenilin 1 Antibody (CAB19103)	PSEN1 (Phospho-Ser357) Colorimetric Cell-Based ELISA Kit
Anti-PSEN1 Antibody (CAB2187)
PSEN1 Antibody (PACO03027)
Phospho-PSEN1 (S357) Antibody (PACO04777)
PSEN1 Antibody (PACO04778)