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TAMRA-(BETA)-amyloid (1-42)

Go to "Antibodies for Neurodegenerative Disease Research"

Background

This product is β-amyloid labelled N-terminus with fluorophore Carboxytetramethylrhodamine (TAMRA). Amyloid-beta (Aβ) is peptides of 36 - 43 amino acids that are crucially involved in Alzheimer's disease as the main component of theamyloid plaques found in the brains of Alzheimer patients. It is useful for research of neurodegenerative disease such as Alzheimer's disease.

Kit component

Components
Product Name Size Quantity Storage Conditions Stability
TAMRA-β-amyloid (1-42) 1.0mg/tube 1 -20°C and protect from light 2 year

[Materials required but not provided]
・ Dimethyl sulfoxide (DMSO) ・ DMEM (phenol red free)
・ Fluorescence microscope

Precautions
1. Read the instructions carefully before use.
2. Wear eye protectors and gloves

Protocol

A) For the unaggregation protocol

  1. Amyloid peptide is dissolved in 200µL of HFIP and is incubated RT until clear solution.
  2. Allow HFIP to evaporate under vacuum by using SpeedVac until the peptide film is observed at the bottom. Store peptide films at –20°C. The peptide film can be stored for several months to years with N2 gas sealing. If the storage period is over several months, need to do again from the first step.
  3. Resolved peptide film by adding 80µL of DMSO to a concentration of 2.5mM and vortex for 30 seconds. Peptides in solution can be stored at -20°C for 2 weeks protected from light.
  4. Dilute the peptide solution to 0.01-10µM with warmed culture medium.
  5. Add the medium including peptide on the cell and culture at 37°C for 24h.
  6. Remove the supernatant, wash with buffer and add the fresh medium.
  7. Observe by fluorescence microscope.

B) For the oligomers protocol

  1. Amyloid peptide is dissolved in 200µL of HFIP and is incubated RT until clear solution.
  2. Allow HFIP to evaporate under vacuum by using SpeedVac until the peptide film is observed at the bottom. Store peptide films at –20°C. The peptide film can be stored for several months to years with N2 gas sealing. If the storage period is over several months, need to do again from the first step.
  3. Resolved peptide film by adding 80µL of DMSO to a concentration of 2.5mM and vortex for 30 seconds.
  4. Dilute the peptide solution to 1:25 with DMEM (phenol red free) to a concentration of 100µM.
  5. Incubate at 4°C for 24h to oligomerize. Oligomerized peptide can be stored at 4°C for 4 weeks protected from light.
  6. Dilute the oligomerized peptide to 0.01-10µM with culture medium.
  7. Add the medium including oligomerized peptide on the cell and culture at 37°C for 24h.
  8. Remove the supernatant, wash with buffer and add the fresh medium.
  9. Observe by fluorescence microscope.

Application Example

Phagocytosed TAMRA-β-amyloid by microglia cell clone

Confocal fluorescent images showing microglia cell clone(Ra2) incorpolated oligomerized TAMRA-β-amyloid.

Product List

Product Name Cat# Quantity Price

TAMRA-(BETA)-amyloid (1-42)

PMC-AK13A-COS 1MG

¥ 50,000
$ 667
€ 500

References
  • Dahlgren, Karie N, Arlene M Manelli, W Blaine Stine, Lorinda K Baker, Grant a Krafft, and Mary Jo LaDu,‘Oligomeric and Fibrillar Species of Amyloid-beta Peptides Differentially Affect Neuronal Viability.’, The Journal of biological chemistry, 277(2002), 32046.53

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.