Figure 1: α-Synuclein protein is detected in mouse or rat brain tissues using the PathScan® Total α-Synuclein Sandwich ELISA Kit. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blot using α-Synuclein (D37A6) XP® Rabbit mAb #4179 is shown in the bottom figure.
Figure 2: The relationship between protein concentration of lysates from extracts of mouse brain and the absorbance at 450 nm as detected by PathScan® Total α-Synuclein Sandwich ELISA Kit is shown. Mouse brains were frozen, ground, and then lysed with lysis buffer #9803.
|Product Includes||Volume||Solution Color|
|α-Synuclein Rabbit mAb Coated Microwells||96 tests|
|α-Synuclein Mouse Detection mAb||1 ea||Green (Lyophilized)|
|Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)||1 ea||Red (Lyophilized)|
|Detection Antibody Diluent||11 ml||Green|
|HRP Diluent||11 ml||Red|
|TMB Substrate 7004||11 ml||Colorless|
|STOP Solution 7002||11 ml||Colorless|
|ELISA Wash Buffer (20X) 9801||10 ml|
|Sealing Tape||2 ea|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) 9803||15 ml||Yellowish|
The PathScan® Total α-Synuclein Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of
α-Synuclein protein. An α-Synuclein rabbit mAb has been coated onto the microwells. After incubation with cell lysates, the α-Synuclein protein is captured by the coated antibody. Following extensive washing, α-Synuclein mouse detection mAb is added to detect captured α-Synuclein protein. Anti-mouse, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for the developed color is proportional to the quantity of α-Synuclein protein. Antibodies in the kit are custom formulations specific to the kit.
NOTE: Prepare solutions with purified water.
*NOTE: Some PathScan® ELISA Kits may include HRP-Linked Streptavidin in place of HRP-Linked Antibody.
NOTE: Initial color of positive reaction is blue, which changes to yellow upon addition of STOP Solution.
posted November 2013
Protocol Id: 204
PathScan® Total α-Synuclein Sandwich ELISA Kit detects endogenous levels of α-Synuclein protein in mouse or rat tissues, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
α-Synuclein is a protein of 140-amino acids expressed abundantly in the brain. α-Synuclein is also the main component of pathogenic Lewy bodies and Lewy neurites. Research studies have shown that mutations of the α-synuclein gene are linked to Parkinson's disease (1).
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