Apoptosis Inhibitor of Macrophage (hAIM) ELISA kit - Cosmo Bio Co.,Ltd.

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Apoptosis Inhibitor of Macrophage (hAIM) ELISA kit

Catalog No.: KAL-KK901
Size: 1KIT
Price: ¥125000
$1667
antigen/source: Apoptosis Inhibitor of Macrophage (hAIM
Application: Enzyme Linked Immunosorbent Assay
Storage: 4C
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Immunogen: Human
Reacts with: Human
Purpose: For accurate measurement of AIM
component: › Antibody Coated Wells --- 1 (6 strips)
› Biotinylated Antibody (30~) --- 1 (200 uL)
› HRP-labeld Avidin (30~) --- 1 (200 uL)
› Enzyme Diluent --- 1 (12 mL)
› Standard/Sample Diluent --- 1 (30 mL)
› Wash Concentrate (20~) --- 1 (30 mL)
› TMB Substrate Reagent --- 1 (12 mL)
› Stop Solution (sulfuric acid) --- 1 (12 mL)
› hAIM Standard (freeze dried) --- 1
(add 250uL distilled water just before use)
MATERIALS REQUIRED BUT NOT PROVIDED
› Microplate reader capable of measurement at 450 nm [reference wavelength at around 650 nm]
› Precision pipettes or multichannel pipettes to deliver 10 uL -1000 uL volumes, and disposable tips
› Tubes with low-protein absorbent to prepare standard dilutions
› Measuring cylinder or beaker (500 mL) to prepare wash solution
› Distilled water
OPERATING PRECAUTIONS
› Do not mix reagents from different kit lots.
› Diluted reagents should not be stored for more than 24 hours.
› Do not discontinue the operation during the experimentation. Also, do not leave the microplate dehydrated during operation.
› Use microplate lid to prevent drying during operation if needed.
› All standards and samples are recommended to be run in duplicate.

Other:

 Background 
AIM (Apoptosis Inhibitor of Macrophage), a protein produced by macrophage and present in the blood, interacts with both adipose cells and macrophages) . By inducing neutral fat breakdown in adipose cells and inhibits cellular death ofmacrophage, AIM shows a strong association with various lifestyle-related diseases such as obesity, arteriosclerosis, and diabetes2-5). In recent years, it is indicated that AIM plays a role in the pathogenesis of autoimmune disease associated with obesity 6). As seen above, amount of AIM in the blood is related to pathological condition of many diseases problematic in modern society, and the measurement of AIM is suggested to be effective for diagnostic and therapeutic research of the lifestyle-related diseases. Precise measurement may be difficult depending on the antibody character since AIM is attached to IgM in the blood, however the antibody used in this kit is unaffected by IgM, which enables accurate measurement of AIM7).
This assay kit employs sandwich ELISA method utilizing mouse monoclonal antibody specific for human AIM (hAIM) protein. hAIM-specific antibody is pre-coated on the microplate. hAIM in standard or specimen (biological sample such as cell culture supernatantand serum) binds specifically to antibody attached on each well of microplate. Then, biotinylated hAIM-specific antibody and HRP-labeled Avidin are added. Finally, colorimetric substrate (TMB) is added and reacted, which produces a color in proportionto the amount of hAIM in the sample. After stopping the enzyme reaction by adding sulfuric acid, the microwell absorbances are measured for the dominant wavelength at 450 nm (reference wavelength: around 650 nm) utilizing microplate reader. Plotthecalibration curve using hAIM Standard, and hAIM concentration in the specimen is determined by finding the absorbance value on the calibration curve.



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Reference:

•  Miyazaki. T. et al., (1999) J. Exp. Med 189: 413-422.
•  Arai, S.et al., (2005) Cell Metab. 1: 201-213.
•  Kurokawa, J.et al., (2010) Cell Metab. 11: 479-492.
•  Kurokawa, J.et al., (2011) Proc. Natl. Acad. Sci. USA. 108: 12072-12077.
•  Miyazaki,T.et al., (2011) Cir. J. 75: 2522-2531.
•  Arai, S.,et al., (2013) Cell Rep., 3: 1187-98.
•  Oba, M.et al., (2012) Seikagaku 84: 588-591.
 
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