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Purity: =99.66%
Amyloid β-Peptide (1-42) human is a human form of amyloid β-peptide composed of 42-amino acids and can be found in the brains of patients with Alzheimer's disease. It plays a key role during the pathogenesis of Alzheimer disease (AD).
| Targets |
Peptide; AD biomarker
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|---|---|
| ln Vitro |
Guidelines for beta-amyloid aggregation (The following are our recommended protocols. This is a guide only and may be modified to suit your specific needs).
1. Dissolve solid Aβ peptide in cold hexafluoro-2-propanol (HFIP). The peptide was incubated at room temperature for at least 1 hour to establish monomers and structural randomization. 2. HFIP is removed by evaporation and the resulting peptide is stored in the form of a thin film at -20 or -80℃. 3. Dissolve the resulting film in 5mM anhydrous DMSO, and then vortex and dilute to the appropriate concentration and buffer (serum and phenol red free medium). 4. Next, leave the solution at 4-8°C for 48 hours. The sample was then centrifuged at 14000g at 4-8°C for 10 minutes; Soluble oligomers in the supernatant. Dilute the supernatant by 10-200 times before the experiment. Methods vary depending on downstream application. Note: The aggregated form is unstable in solution and is recommended for immediate use. |
| ln Vivo |
Alzheimer's disease models in animals can be created using human TFA and β-Amyloid (1-42).
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| Enzyme Assay |
The possibility to monitor, in solution, the steps of beta-amyloid (Abeta) nucleation and therefore to describe this dynamic process by using capillary electrophoresis and under optimized experimental conditions is described. Striking differences in the electrophoretic patterns of Abeta 1-42 and Abeta 1-40 over time are here shown, and different aggregation states are elucidated, which reflect the very diverse oligomerization behavior of two very similar peptides. The isolation of one aggregated species of high molecular weight by ultracentrifugation allowed us to assess its role as toxic oligomer. The perturbation of the existing equilibrium among the identified species by the addition of small molecules can in principle interfere with the aggregation process of the peptides and ultimately prevent the plaque formation in vitro.[3]
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| Cell Assay |
Different types of voltage-gated ion currents were recorded in isolated neurons of snail Helix pomatia using the two-microelectrode voltage-clamp technique. Application of amyloid-β peptide (1-42, 1-10 μM) in the bathing solution did not change delayed rectifier K(+)-current and leakage current, but enhanced inactivation of Ca(2+)-current and blocked Ca(2+)-dependent K(+)-current.[1]
Aβ Peptides and MTT Assay [2] Synthetic Aβ peptides were monomerized and solubilized as described. Briefly, monomerized peptides were dissolved to 1 mg/ml in deionized water supplemented with ammonia to a final concentration of 0.13% (measured at pH 9.8). All peptides were used at a concentration of 1 μm. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed as described previously. Nuclei Preparation and ELISA[2] Nuclei were isolated with the Nuclei EZ Prep nuclei isolation kit according to the manufacturer's instructions with minor modifications. Briefly, pellets containing nuclei were resuspended in PBS, sonicated, incubated at 95 °C, and centrifuged for 10 min at 20,000 × g. Aβ42 ELISAs using the C-terminal specific G2-13 antibody were performed as described. To quantify Aβ38, Aβ40, Aβ42, Aβ42 G33A, and Aβ43, the antibody 4G8 recognizing Aβ residues 17–24 was used. Immunofluorescence [2] SH-SY5Y cells were treated with biotinylated Aβ42 peptide 1 μm. Cells were fixed and permeabilized with 3.3% formaldehyde containing 0.5% Triton X-100 followed by 125 mm glycine in PBS containing magnesium and calcium. Cells were blocked with 5% fetal bovine calf serum followed by the primary antibody. Biotin-Aβ42 was detected with the monoclonal antibody AB or alternatively with Avidin Fluor488 (Sigma). Nuclei were stained with DAPI. Images were obtained using an LSM 510 meta confocal microscope. |
| Animal Protocol |
Animals (transgenic APPPS1 mice, 12 months old) were perfused with PBS followed by fixative solution (4% formaldehyde, 0.2% glutaraldehyde in 50 mm sodium cacodylate buffer, pH 7.4). Hippocampi were dissected and post-fixed in the same solution at room temperature. Samples were embedded in LR-Gold resin and polymerized at 4 °C. Ultra-thin sections were incubated with the G2-13 primary antibody labeled with 10 nm colloidal gold. The sections were counterstained with uranyl acetate followed by lead citrate.[2]
Labeling of mAb G2-13 was performed with colloidal gold. After centrifugation (10 min, 6700 × g), the supernatant of colloidal gold was counterstained with uranyl acetate and analyzed by TEM to ensure that the supernatant was free of gold aggregates. An amount of 500 μg of mAb G2-13 was dialyzed with 2 mm sodium tetraborate. Equal volumes of G2-13 and colloidal gold were incubated for 20 min at RT. The stability of the gold/protein ratio was assessed by titration with 10% NaCl. Colloidal gold was adjusted to pH 9 in 100 mm potassium carbonate. The protein/gold solution was incubated in 1% BSA for 20 min. After centrifugation, the pellet was resuspended in 20 mm TBS, 1% BSA, and 0.05% sodium azide, pH 8.2, and the solution was centrifuged for 5 min at 6700 × g. The supernatant contained G2-13 antibodies labeled with 10 nm of gold.[2] |
| References |
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| Additional Infomation |
Different types of voltage-gated ion currents were recorded in isolated neurons of snail Helix pomatia using the two-microelectrode voltage-clamp technique. Application of amyloid-β peptide (1-42, 1-10 μM) in the bathing solution did not change delayed rectifier K(+)-current and leakage current, but enhanced inactivation of Ca(2+)-current and blocked Ca(2+)-dependent K(+)-current.[1]
Although soluble species of the amyloid-β peptide Aβ42 correlate with disease symptoms in Alzheimer disease, little is known about the biological activities of amyloid-β (Aβ). Here, we show that Aβ peptides varying in lengths from 38 to 43 amino acids are internalized by cultured neuroblastoma cells and can be found in the nucleus. By three independent methods, we demonstrate direct detection of nuclear Aβ42 as follows: (i) biochemical analysis of nuclear fractions; (ii) detection of biotin-labeled Aβ in living cells by confocal laser scanning microscopy; and (iii) transmission electron microscopy of Aβ in cultured cells, as well as brain tissue of wild-type and transgenic APPPS1 mice (overexpression of amyloid precursor protein and presenilin 1 with Swedish and L166P mutations, respectively). Also, this study details a novel role for Aβ42 in nuclear signaling, distinct from the amyloid precursor protein intracellular domain. Chromatin immunoprecipitation showed that Aβ42 specifically interacts as a repressor of gene transcription with LRP1 and KAI1 promoters. By quantitative RT-PCR, we confirmed that mRNA levels of the examined candidate genes were exclusively decreased by the potentially neurotoxic Aβ42 wild-type peptide. Shorter peptides (Aβ38 or Aβ40) and other longer peptides (nontoxic Aβ42 G33A substitution or Aβ43) did not affect mRNA levels. Overall, our data indicate that the nuclear translocation of Aβ42 impacts gene regulation, and deleterious effects of Aβ42 in Alzheimer disease pathogenesis may be influenced by altering the expression profiles of disease-modifying genes.[2] |
| Molecular Formula |
C₂₀₃H₃₁₁N₅₅O₆₀S
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|---|---|
| Molecular Weight |
4514.04
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| Exact Mass |
4511.27
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| CAS # |
107761-42-2
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| Sequence |
Asp-Ala-Glu-Phe-Arg-His-Asp-Ser-Gly-Tyr-Glu-Val-His-His-Gln-Lys-Leu-Val-Phe-Phe-Ala-Glu-Asp-Val-Gly-Ser-Asn-Lys-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala
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| SequenceShortening |
DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA
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| Appearance |
White to off-white solid powder
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| LogP |
1.351
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| Chemical Name |
(4S)-5-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[2-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-5-amino-1-[[(2S)-6-amino-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[2-[[(2S)-1-[[(2S)-4-amino-1-[[(2S)-6-amino-1-[[2-[[(2S)-1-[[(2S,3S)-1-[[(2S,3S)-1-[[2-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[2-[[2-[[(2S)-1-[[(2S)-1-[[(2S,3S)-1-[[(1S)-1-carboxyethyl]amino]-3-methyl-1-oxopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-2-oxoethyl]amino]-2-oxoethyl]amino]-3-methyl-1-oxobutan-2-yl]amino]-4-methylsulfanyl-1-oxobutan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-2-oxoethyl]amino]-3-methyl-1-oxopentan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-1-oxohexan-2-yl]amino]-1,4-dioxobutan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-3-methyl-1-oxobutan-2-yl]amino]-3-carboxy-1-oxopropan-2-yl]amino]-4-carboxy-1-oxobutan-2-yl]amino]-1-oxopropan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1-oxohexan-2-yl]amino]-1,5-dioxopentan-2-yl]amino]-3-(1H-imidazol-4-yl)-1-oxopropan-2-yl]amino]-3-(1H-imidazol-4-yl)-1-oxopropan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-4-carboxy-1-oxobutan-2-yl]amino]-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-3-carboxy-1-oxopropan-2-yl]amino]-3-(1H-imidazol-4-yl)-1-oxopropan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-4-[[(2S)-2-[[(2S)-2-amino-3-carboxypropanoyl]amino]propanoyl]amino]-5-oxopentanoic acid
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| Synonyms |
β-Amyloid (1-42, human
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: (1). This product is not stable in solution, please use freshly prepared working solution for optimal results. (2). Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~33.33 mg/mL (~7.20 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (0.54 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 2.5 mg/mL (0.54 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (0.54 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.2215 mL | 1.1077 mL | 2.2153 mL | |
| 5 mM | 0.0443 mL | 0.2215 mL | 0.4431 mL | |
| 10 mM | 0.0222 mL | 0.1108 mL | 0.2215 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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