1. Alzheimer's Disease Research

for quantitative determination of βAmyloid peptide 40 and 42
β Amyloid ELISA Kits
Alzheimer's Disease (AD) is characterized by the presence of extracellular senile plaques (SPs) and intracellular neurofibrillary tangles (NFT) in the brain. The major protein component of SPs is β Amyloid peptide (Aβ) 40 and 42(43). Aβ42 is more prone to aggregate than Aβ. Therefore the initial Abdeposition begins with Aβ42(43) but not with Aβ40. Aβ42(43)-positive and Aβ40-negative plaques may represent early-stage diffuse type SPs, and Aβ40-positive plaque appears in the advanced stage, especially more often in the cored portion of the mature plaque.
In these kits, we use the monoclonal antibodies which specifically detect Aβ. Therefore these kits are designed to be used for the quantitative determination of Aβ in samples such as tissue culture medium, tissue homogenate, CSF and plasma.
[Features]
1. These kits are designed to be used for the quantitative determination of Aβ in samples such as tissue culture medium, tissue homogenate, CSF and plasma.
2. These kits use the monoclonal antibodies that were developed by Takeda Pharmaceutical Company, Ltd.
BAN50: specifically detects the N-terminal of Aβ (1-16)
BNT77: specifically detects the Aβ (11-28) of Aβ
BA27: specifically detects the C-terminal of Aβ 40
BC05: specifically detects the C-terminal of Aβ 42
[Kit Contents] ( Keep at 2~10 °C)
    1) MAb-coated Microtiter Plate1 plate
    2) Standard Solution2 vials x 2 mL
    3) Standard Diluent1 vial x 30 mL
    4) Wash Solution (20 x)1 vial x 50 mL
    5) HRP-conjugated MAb Solution1 vial x 12 mL
    6) TMB Solution1 vial x 12 mL
    7) Stop Solution1 vial x 12 mL
    8) Plate Seal3 sheets
Click here to see the detailed information
    ⇒ Detailed information on the assay (HTML)



*: Improved β Amyloid (1-40) and (x-40)*** Detection Kits
These products are detection kits that achieve more stable antigenantibody reactions by using F(ab')2 fragment of the labeled antibody BA27, which recognizes the C-terminal of Aβ40, while the nonspecific binding is remaining to be reduced. Therefore, the stability in washing solution is improved. In Human/Rat kit, background is reduced.
**: Highly Sensitive β Amyloid (1-42) and (x-42)*** Detection Kits
These ELISA kits detecting β amyloid (1-42) and (x-42) that ensure highly sensitive detection. These β amyloids are indicated to have a correlation with Alzheimer's disease. Detection sensitivity of the kits is about 10 times higher than that of the conventional products and the dynamic range is from 0.1 to 20.0 pmol/L. Fab' fragment of the labeled antibody is used as well so that the nonspecific binding is reduced.
*** Ab (x-40) and (x-42) are Aβ peptide modifi ed or cleaved at the N-terminal.

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Standard Curve

Human β Amyloid (1-42) ELISA Kit Wako, High-Sensitive ()
Human/Rat β Amyloid (42) ELISA Kit Wako, High-Sensitive ()
Human β Amyloid (1-40) ELISA Kit Wako II()
Human/Rat β Amyloid (40) ELISA Kit Wako II ()

Reaction time for HRP-conjugated antibody is 2 hours.

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Application Data [1] human and mouse plasma

Blood was collected using vacuum blood collection tube containing EDTA-2K. Plasma was separated by centrifugation at 5000 x g for 15 minutes at 4°C and stored at -80°C until used.
The plasma sample was diluted 4-fold with Standard Diluent in the Kit and measured. (Aβ(1-40) and Aβ (x-40) were measured by HumanAmyloid (1-40) ELISA Kit Wako II (Cat.#298-64601, ) and Human/Rat β Amyloid (40) ELISA Kit Wako II (Cat.#294-64701, ))
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Application Data [2] mouse brain tissue

The hemisphere of 12-month mouse (J20) was extracted with 2 mL of Tris Saline and stored frozen at -20°C until used. The brain sample was diluted 2-fold with Standard Diluent in the Kit and measured. Aβ trace quantity of A could be detected not only in the transgenic mice (tg) but in the wildtype mice (wt).
(Data provided by Prof. Iwatsubo and Instructor Hashimoto, Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, University of Tokyo)
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Specificity (n=4) , Spike Recovery (n=3)

Human β Amyloid (1-42) ELISA Kit, High-Sensitive(3) / Human / Rat B Amyloid (42) ELISA Kit, High-Sensitive(6)

II Kit-conventional kit Correlation


[References]
1) Suzuki N., Cheung TT., Cai XD., Odaka A., Otvos L. Jr., Eckman C., Golde TE. And Younkin SG: Science, 264, 1336 (1994).
2) Iwatsubo T., Odaka A., Suzuki N., Mizusawa N. and Ihara Y.: Neuron, 13, 45 (1994).
3) Asami-Odaka A., Ishibashi, Y., Kikuchi T., Kitada C. and Suzuki N.: Biochemistry, 34, 10272 (1995).
4) Fukumoto H., Tomita T., Matsunaga H., Ishibashi Y., Saido T.C. and Iwatsubo T.: Neuroreport, 10, 2965 (1999).
5) Scheuner D., Eckman C., Jensen M., Song X., Citron M., Suzuki N., Bird TD., Hardy J., Hutton M., Kukull W., Larson E., etc.: Nature Med., 2, 864 (1996).
6) Kosaka T., Imagawa M., Seki K., Arai H., Sasaki H., Tsuji S., Asami-Odaka A., Fukushima T., Imai K. and Iwatsubo T.: Neurology, 48, 741 (1997).
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Histostaining of Alzheimer's Diseased Brain Tissues
    - Distinctive Histostaining of Aβ40 and Aβ42 plaques
Amyloid β-Protein Immunohistostain Kit
image
Immunostaining of senile plaques in the consecutive sections of the brain affected with Alzheimer's Disease.
Left : Aβ42-staining using Anti Aβ42 Ab (Clone #BC05);
Right : Aβ40- staining using Anti Aβ40 Ab (Clone #BA27)

(provided by Dr. Iwatsubo, Univ. of Tokyo)

Wako Cat. No. 299-56701 (50 tests)
Keep at 2-10°C
[Features]
1. Distinctive histostaining of Aβ40 and 42 plaques in Alzheimer's diseased brain tissues
2. High sensitivity histo-Immune detection of Aβ plaques in tissue sections with low background

[Kit Contents (50 tests)]
1) Blocking Serum1 bottle x 10 mL
2) Anti Mouse lgG (H+L), Goat, Conjugated1 bottle x 10 mL
3) ABC Solution
     (Streptavidin-biotin-peroxidase Complex)
1 bottle x 10 mL
4) Formic Acid (90%) 1 bottle x 15 mL
5) Anti Amyloid β-Protein (1-40), MAb,
     Clone # BA27
1 bottle x 7 mL
6) Anti Amyloid β-Protein (1-42), MAb,
     Clone # BV05
1 bottle x 7 mL
7) Trypsin, Cryst 1 bottle x 50 mL
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Wako Product Update Bio-No.1 [ page. 3-5 ]