Figure 1. The PathScan® β-Amyloid (1-43) Sandwich ELISA Kit #43825 detects human Aβ-43 and to a lesser degree Aβ-42 but not Aβ-40, Aβ-39, Aβ-38, Aβ-37, and pE3 peptides. The absorbance at 450 nm using the PathScan® β-Amyloid (1-43) Sandwich ELISA Kit #43825 for a panel of human β-amyloid peptides is shown in the figure.
Figure 2. The relationship between the concentration of human Aβ-43, Aβ-42 or Aβ-40 peptide and the absorbance at 450 nm using the PathScan® β-Amyloid (1-43) Sandwich ELISA Kit #43825 is shown.
|Product Includes||Volume||Solution Color|
|β-Amyloid (1-43) Rabbit mAb Coated Microwells||96 tests|
|β-Amyloid Mouse Detection mAb||1 ea||Red (Lyophilized)|
|HRP Diluent||5.5 ml||Red|
|TMB Substrate 7004||11 ml|
|STOP Solution 7002||11 ml|
|Sealing Tape||2 ea|
|ELISA Wash Buffer (20X) 9801||25 ml|
|Cell Lysis Buffer (10X) 9803||15 ml|
The PathScan® β-Amyloid (1-43) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of the Aβ-43 isoform of β-amyloid. This kit also detects to a lesser extent Aβ-42, as shown in Figures 1 and 2. Incubation of cell lysates and detection antibody on the coated microwell plate forms a sandwich with Aβ-43 in a single step. The plate is then extensively washed and TMB reagent is added for signal development. The magnitude of absorbance for the developed color is proportional to the quantity of Aβ-43.
*Antibodies in this kit are custom formulations specific to kit.
NOTE: This protocol is for PathScan® kits that use an HRP directly conjugated to the detection antibody (1-step method), rather than a 2-step method where the detection antibody and a secondary-HRP are added sequentially.
Refer to product-specific datasheets for assay incubation temperature.
NOTE: Prepare solutions with deionized/purified water or equivalent.
For adherent cells
For suspension cells
NOTE: Equilibrate all materials and prepared reagents to room temperature prior to running the assay.
NOTE: Initial color of positive reaction is blue, which changes to yellow upon addition of STOP Solution.
created July 2020
Protocol Id: 2144
The PathScan® β-Amyloid (1-43) Sandwich ELISA Kit detects endogenous levels of the Aβ-43 isoform of β-amyloid peptides, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit also detects to a lesser extent Aβ-42, as shown in Figures 1 and 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Amyloid β (Aβ) precursor protein (APP) is a 100-140 kDa transmembrane glycoprotein that exists as several isoforms (1). The amino acid sequence of APP contains the amyloid domain, which can be released by a two-step proteolytic cleavage (1). The extracellular deposition and accumulation of the released Aβ fragments form the main components of amyloid plaques in Alzheimer's disease (1). APP can be phosphorylated at several sites, which may affect the proteolytic processing and secretion of this protein (2-5). Phosphorylation at Thr668 (a position corresponding to the APP695 isoform) by cyclin-dependent kinase is cell-cycle dependent and peaks during G2/M phase (4). APP phosphorylated at Thr668 exists in adult rat brain and correlates with cultured neuronal differentiation (5,6).
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