Western blot of purified anti-β-amyloid, 1-16 antibody (clone 6E10). Lane 1: Molecular weight marker; Lane 2: 50 ng of the human Aβ1-40 peptide; Lane 3: 50 ng of the Aβ1-42 peptide; Lane 4: 50 ng of the recombinant human APP751 protein; Lane 5: 20µg of the human brain lysate; Lane 6: 50 ng of the rodent Aβ1-42 peptide. The blot was incubated with 1 µg/mL of the primary antibody overnight at 4°C, followed by incubation with the HRP goat anti-mouse IgG antibody (Cat. No. 405306). Enhanced chemiluminescence was used as the detection system.
Western blot of purified anti-β-amyloid, 1-16 antibody (clone 6E10). Lane 1: Molecular weight marker; Lane 2: 50 ng of the human Aβ1-40 peptide; Lane 3: 50 ng of the Aβ1-42 peptide; Lane 4: 50 ng of the recombinant human APP751 protein; Lane 5: 20µg of the human brain lysate; Lane 6: 50 ng of the rodent Aβ1-42 peptide. The blot was incubated with 1 µg/mL of the primary antibody overnight at 4°C, followed by incubation with the HRP goat anti-mouse IgG antibody (Cat. No. 405306). Enhanced chemiluminescence was used as the detection system.
Direct ELISA of purified anti-β-amyloid 1-16 (clone 6E10) antibody binding to the plate-immobilized human Aβ1-40 and the recombinant human APP751. ELISA was performed by coating the wells with 100 ng of peptide or recombinant protein. The wells were then incubated with the primary antibody at 37°C for 45 minutes, followed by incubation with HRP labeled goat anti-mouse IgG secondary antibody. TMB (3, 3', 5, 5' tetramethylbenzidine, Cat. No. 421501) was used as the detection system.
IHC staining of purified anti-β-Amyloid, 1-16 antibody (clone 6E10) on the formalin-fixed paraffin-embedded normal human (left panel) and Alzheimer’s disease (right panel) brain tissues. Following antigen retrieval using 70% formic acid for 20 minutes, the tissues were incubated with 1µg/mL of the primary antibody for 1 hour at room temperature. BioLegend’s Ultra-Streptavidin (USA) HRP Detection Kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The images were captured with a 40X objective. Scale bar: 50 µm
Alzheimer's disease is characterized by the accumulation of aggregated Aβ peptides in senile plaques and vascular deposits. Aβ peptides are derived from amyloid precursor proteins (APP) through sequential proteolytic cleavage of APP by β-secretases and γ-secretases generating diverse Aβ species. Aβ can aggregate to form soluble oligomeric species and insoluble fibrillar or amorphous assemblies. Some forms of the aggregated peptides are toxic to neurons.
Phosphate-buffered solution (no preservatives or carrier proteins).
Preparation
The antibody was purified by affinity chromatography.
Concentration
1 mg/mL
Storage & Handling
This antibody should be handled aseptically as it is free of preservatives such as Sodium Azide. Store this antibody undiluted between 2°C and 8°C. Please note the storage condition for this antibody has been changed from -20°C to between 2°C and 8°C. You can also check the vial label or CoA to find the proper storage conditions.
Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 1 µg/mL. For Direct ELISA, the suggested use of this reagent is 0.002 - 0.02 µg/mL. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, the suggested use of this reagent is 0.2 - 5.0 µg/mL. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes
This antibody is reactive to amino acid residue 1-16 of beta amyloid. The epitope lies within amino acids 3-8 of beta amyloid (EFRHDS).
This antibody clone has been reported for use in immunohistochemistry of free-floating sections2,13.
Amyloid precursor protein is a 770 amino acid protein with a molecular mass of ~100 kD. According to the UniProtKB database, APP (ID# P05067) has 11 isoforms (34 to ~90 kD) and the 770 form has been designated as the canonical form. Isoform APP695 is the predominant form expressed in neuronal tissue. Isoforms APP751 and APP770 are widely expressed in non-neuronal cells. Isoform APP751 is the most abundant form in T-lymphocytes. Aβ denotes peptides of 36-43 amino acids generated from cleavage of APP by secretases. Aβ has an apparent molecular mass of about 4 kD.
Distribution
Tissue distribution: Primarily nervous system, but also adipose tissue, intestine, muscle.
Cellular distribution: Cytosol, endosomes, nucleus, plasma membrane, extracellular, and golgi apparatus.
Function
The normal function of Aβ is not well understood. Several potential physiological roles have been proposed, including: activation of kinase enzymes; protection against oxidative stress; regulation of cholesterol transport; transcription factor, and as an anti-microbial agent.
Biology Area
Cell Biology, Neurodegeneration, Neuroscience, Protein Misfolding and Aggregation
Molecular Family
APP/β-Amyloid
Antigen References
Kumar A, et al. 2015. Pharmacol. Rep. 67(2):195.
Sadigh-Eteghad S, et al. 2015. Med. Princ. Pract. 24(1):1
Hampel H, et al. 2015. Expert Rev. Neurother. 15(1):83.
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