Product DescriptiongoogleSynthetic beta-amyloid Aβ1-42 was monomerized by HFIP (hexafluoro-2-propanol) treatment and dried. One vial contains 50 μg monomeric Aβ peptide that can be used to form solutions of unaggregated Aβ monomers, aggregated Aβ oligomers, Aβ fibrils and Aβ protein complexes according to published protocols, and used in a variety of research applications.
Alternative NamesBeta-APP42; Beta-amyloid protein 42; ABPP; APPI; Amyloid beta A4 protein; AB42; abeta
Product DescriptionSynthetic beta-amyloid Aβ1-42 was monomerized by HFIP (hexafluoro-2-propanol) treatment and dried. One vial contains 50 μg monomeric Aβ peptide that can be used to form solutions of unaggregated Aβ monomers, aggregated Aβ oligomers, Aβ fibrils and Aβ protein complexes according to published protocols, and used in a variety of research applications.
Application DetailsPreparation of unaggregated A-beta1-42:
Important: unaggregated A-beta has to be prepared just prior to use!
1. Add 5 µL of reconstituting buffer to one vial of 50 µg of HFIP-treated A-beta peptide; spin down the liquid briefly 2. Vortex the vial for 5 seconds at highest speed while rotating the vial with your hands; spin down the liquid (bench-top microcentrifuge) and repeat the vortex-spin procedure for a minimum of 3 times; continue the vortex-spin procedure until all lyophilized peptide is dissolved and collected at the bottom of the tube. Important: refer to the attached instructions for a detailed procedure to ensure that all peptide is fully reconstituted! 3. Add 106 µL of cold Dilution Buffer to make up to 111 µL total volume and a peptide concentration of 100 µM. Vortex-spin for 3 more times 4. Final concentration of A-beta is 450 µg/mL 5. Use reconstituted peptide immediately to avoid oligomer formation
Preparation of oligomeric A-beta1-42:
1. Add 5 µL of reconstituting buffer to one vial of 50 µg of HFIP-treated A-beta peptide; spin down the liquid briefly 2. Vortex the vial for 5 seconds at highest speed while rotating the vial with your hands; spin down the liquid (bench-top microcentrifuge) and repeat the vortex-spin procedure for a minimum of 3 times; continue the vortex-spin procedure until all lyophilized peptide is dissolved and collected at the bottom of the tube. Important: refer to the attached instructions for a detailed procedure to ensure that all peptide is fully reconstituted! 3. Add 106 µL of cold Dilution Buffer to make up to 111 µL total volume and a peptide concentration of 100 µM 4. Vortex-spin for 3 more times 5. Incubate the solution at 2-8ºC for 24 hours (protected from light) 6. Final concentration of A-beta is 450 µg/mL 7. Once reconstituted and oligomerized, o-A-beta should be used as soon as possible and within 7 days to ensure the stability of the oligomers
Note: while the concentration of monomeric A-beta peptide used to form the oligomeric complexes is accurately determined, the precise formation, size and number of oligomers cannot be quantified by any known method.
Preparation of fibrillar A-beta1-42:
1. Add 5 µL of reconstituting buffer to one vial of 50 µg of HFIP-treated A-beta peptide; spin down the liquid briefly 2. Vortex the vial for 5 seconds at highest speed while rotating the vial with your hands; spin down the liquid (bench-top microcentrifuge) and repeat the vortex-spin procedure for a minimum of 3 times; continue the vortex-spin procedure until all lyophilized peptide is dissolved and collected at the bottom of the tube. Important: refer to the attached instructions for a detailed procedure to ensure that all peptide is fully reconstituted! 3. Add 106 µL of 10 mM HCl to make up to 111 µL total volume and a peptide concentration of 100 µM 4. Vortex-spin for 3 more times 5. Incubate the solution at 37ºC for 24 hours (protected from light) 6. Final concentration of A-beta is 450 µg/mL
Preparation of A-beta1-42 Complexes:
Important: only unagreggated A-beta will form complexes. Use A-beta peptide immediately after reconstitution to form complexes.
1. Add 5 µL of reconstituting buffer to one vial of 50 µg of HFIP-treated A-beta peptide; spin down the liquid briefly 2. Vortex the vial for 5 seconds at highest speed while rotating the vial with your hands; spin down the liquid (bench-top microcentrifuge) and repeat the vortex-spin procedure for a minimum of 3 times; continue the vortex-spin procedure until all lyophilized peptide is dissolved and collected at the bottom of the tube. Important: refer to the attached instructions for a detailed procedure to ensure that all peptide is fully reconstituted! 3. Add 106 µL of cold Dilution Buffer to make up to 111 µL total volume and a peptide concentration of 100 µM 4. Vortex-spin for 3 more times 5. Use reconstituted peptide immediately to avoid oligomer formation 6. Mix the A-beta monomer with its complex partner (eg., lipoprotein) at desired concentrations in PBS, pH 7.4, or other suitable buffers compatible with its intended application 7. Incubate at room temperature for 2 hours without shaking 8. Use complexes immediately after incubation
These protocols are based on procedures published by Youmans KL et al., 2012 and Tai LM et al., 2013, and we refer to these publications and other relevant literature for further details.
Provided working concentrations are only meant to guide the user. Optimal concentrations depend on the experimental design and need to be determined empirically.
Kit Components3 vials. 1 x 50 μg Abeta1-42 peptide; 1 x 100 μL Reconstituting Buffer, 1 x 1 mL Dilution Buffer
Purity DescriptionPurified
AppearanceTranslucent film or droplets, dry, without preservatives.
FormatLyophilized.
Reconstitution InstructionsVaries, dependent on intended use. Refer to Applications.
Storage InstructionsStore unopened, dry Aβ peptide vial with desiccant, insulated, at -20°C short term, -80°C long term. Store the buffers at 2-8°C, do not freeze.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Alternative NamesBeta-APP42; Beta-amyloid protein 42; ABPP; APPI; Amyloid beta A4 protein; AB42; abeta
Oligomeric A-beta standard curves generated with BEK-2215 (Oligomeric A beta ELISA kit). Pre-formed oligomers (PE-1750-1000) were reconstituted in assay buffer and compared to oligomers freshly prepared from HFIP-treated A-beta peptide (PE-1749-50). This data demonstrates the usefulness of PE-1749-50 as oligomeric A-beta protein standard.
General ReferencesYoumans KL. et al. (2012) Intraneuronal Abeta detection in 5xFAD mice by a new Abeta-specific antibody. Mol Neurodegener. 2012 Mar 16;7(1):8.
Tai LM. et al. (2013) Levels of Soluble Apolipoprotein E/Amyloid-beta (Aβ) Complex Are Reduced and Oligomeric Abeta Increased with APOE4 and Alzheimer Disease in a Transgenic Mouse Model and Human Samples. J Biol Chem. 2013 Feb 22;288(8):5914-26.