IHC staining of anti-β-Amyloid, 17-24 antibody (clone 4G8) on formalin-fixed paraffin-embedded human Alzheimer's disease brain tissue. Following antigen retrieval using 88% formic acid for 20 minutes at room temperature, the tissue was incubated with the primary antibody at 1:5000 dilution overnight at 4°C. BioLegend´s Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 10X objective. Scale bar: 50 µm
IHC staining of anti-β-Amyloid, 17-24 antibody (clone 4G8) on formalin-fixed paraffin-embedded human Alzheimer's disease brain tissue. Following antigen retrieval using 88% formic acid for 20 minutes at room temperature, the tissue was incubated with the primary antibody at 1:5000 dilution overnight at 4°C. BioLegend´s Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 10X objective. Scale bar: 50 µm
IHC staining of anti-β-Amyloid, 17-24 antibody (clone 4G8) on formalin-fixed paraffin-embedded human Alzheimer's disease brain tissue. Following antigen retrieval using 88% formic acid for 20 minutes at room temperature, the tissue was incubated with the primary antibody at 1:5000 dilution overnight at 4°C. BioLegend´s Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective. Scale bar: 50 µm
Direct ELISA of anti-β-Amyloid, 17-24 (clone 4G8) antibody binding to the plate-immobilized human Aβ1-40, human Aβ1-42 and recombinant human APP751 protein. 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.
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.
The concentration is not quantified as this product is sold as undiluted crude mouse ascites fluid. The concentration might vary from lot-to-lot and an estimated concentration would be 1-3 mg/ml.
Storage & Handling
Store at -20°C or below. Upon initial thawing, apportion into working aliquots and store at -20°C or below. Avoid repeated freeze-thaw cycles to prevent denaturing the antibody. Do not store in frost-free freezers.
Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a dilution range of 1:1000 - 1:5000 is suggested. For Direct ELISA applications, a dilution range of 1:100 - 1:1000 is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes
This antibody is reactive to amino acid residues 17-24 of ß amyloid. The epitope lies within amino acids 18-22 of ß amyloid (VFFAE). 4G8 ß-amyloid antibody reacts to abnormally processed isoforms, as well as precursor forms.
This antibody clone has been reported for use on IHC of free-floating sections in PBS containing 1% Triton incubated with 0.1 m citrate buffer (4).
Application References
(PubMed link indicates BioLegend citation)
Poduslo JF, et al. 2004. Biochem. 43:6064. (IHC-F)PubMed
Forny-Germano L, et al. 2014. J Neurosci. 34:13629. (IHC-Other)PubMed
Vallino Costassa E, et al. 2016. J Alzheimers Dis. 51: 875:87. (IHC-P)PubMed
Chen X, et al. 2013. Neurobiol Aging. 34:2370. (ICC)PubMed
Hatami A, et al. 2016. J Alzheimers Dis. 50:517. (IHC-P)PubMed
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, and 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 Neruother. 15(1):83.
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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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