High Molecular Amyloid β Oligomer ELISA Kit Wako Ver.2

Features

• Detects in serum and plasma, CSF sample
• Specifically reactive with ≧9mer Aβ oligomer

Performance

• Standard curve range	0.41-100 pM (Cerebrospinal fluid) 0.16-40 pM (serum or plasma(EDTA))
  Reactive Aβ oligomers	≧9mer Aβ oligomer
  Sample	Serum, Plasma(EDTA), Cerebrospinal fluid(CSF), In vitro Aβ oligomer
  Cross-reactivity	Human
  Sample volume	CSF:25μL(4 times dilution) Serum, plasma:50μL(2 times dilution)
  Assay time	4.5 hours
  Detection method	Luminescence method**

  • Calculated based on 16-mer MAP peptide.
  • * Requires luminescence plate reader for measurement.

• 

Assay principal

This ELISA kit is designed to measure ≧ 9-mer Aβ oligomer specifically by using same Aβ antibody (BAN50)
for capture and detection³⁾⁴⁾. As a standard, a 16-mer MAP peptide with 16 epitopes of anti Aβ antibody (BAN50) is used⁴⁾.

Aβ monomers were incubated the solution at 4℃ overnight in order to synthesize various size of Aβ oligomers. They were separated Fraction A and Fraction B by ultrafiltration. The fractions were analyzed by western blotting and the Aβ oligomer ELISA.

• Western blotting using Aβ antibody(BAN50)

  

• Aβ oligomer ELISA

  

Example of measurement 1 - spiked recovery test and dilution linearity -

Example of measurement 2 - CSF sample -

Cerebrospinal fluids(CSF) of patients with Alzheimer disease (AD) and Mild Cognitive Impairment (MCI), non-dementia patients(control), were measured with our ELISA kit and competitor’s kit.

• 	Age	Sex	MMSE※3	CSF Aβ oligomer(pM)
  Control n=5	69	M	30	2.37
  	60	F	30	1.65
  	59	M	30	2.82
  	46	M	28	2.24
  	60	F	29	1.34
  MCI n=5	69	F	24	3.6
  	76	M	23	2.62
  	82	M	26	2.33
  	69	F	23	2.67
  	73	F	25	2.34
  AD n=8	79	F	10	3.56
  	88	M	24	3.13
  	62	M	16	4.13
  	78	M	13	3.95
  	63	M	14	3.88
  	74	F	16	3.25
  	76	M	19	3.69
  	84	M	16	4.34

  • ***MMSE score：diagnosis Inquiry test score of cognitive function
    30-28 Normal
    27-24 Suspected of mild cognitive impairment
    ≦23 Suspected of dementia

• 

  • There was a significant difference between Control, MCI and AD.
  • High molecular Aβ oligomer correlated with MMSE score.