Metallothionein (MT) which is Cd binding protein have discovered from horse kidney by Margoshe and Vallee in 1957. MT is present in all animal cells and has strong affinity against another heavy metal.The physiological role are concerned in detoxication of organic metal, metabolic regulation of bioessential metal, protection from stress andfree radical. However, to obtain high affinity anti MT antibody was veryhard because MT is cysteine rich (about 30%), low molecular weight protein and by the absence of aromatic amino acid in this structure.Therefore, the high sensitive ELISA system has not established and difficult to investigate the behavior of MT in vivo.
This kit is high sensitivive ELISA Kit by MT antibody for determining human and animal MT. The epitope of this antibody is located at NH2terminals acetylated peptides in MT, therefore can assay native MT.In addition, NH2 terminals of MT is high conserved sequence in many animal species, therefore almost animals MT can measure in this ELISA kit.
Recently, the Metallothionein (MT) protein is paid to attention in the research of cancer, immunology, the stress, and Alzheimer.
Simultaneous measurement of MTI and MTII.
React with nativw MT. The reaction to expressed MT like non-acetylated N-terminal recombinant protein is 5% or less.
React to various animals such as human, rabbit, mouse, rat, and fishe.
Wide measurement such as blood, tissue, urines, and cell culture supernatant.
|Assay range; 0.7-10000 ng/mL|
|40 samples can be measured with duplicate|
|Microtiter plate is divisible strip, 96wells(12 strips, 8 wells)|
|Sampling volume is 50μL|
|Total reaction time is 2 hrs &10 minutes.|
|Store at 2-8℃.The kit is stable for 6 month after the date of manufacture.|
< Specificity >
The antibody in this kit can recognize the N terminals, especially acetyl group of N terminal amino acid. Therefore, the cross reactivity against recombinat MT protein expressed in bacteria are under 5%.
< Assay principle >
This ELISA kit adopts indirect competitive reaction using the rabbit MT polyclonal antibody which recognize the N terminals of MT. The MT are coated in the surface of 96 wells plate, onto which standard of MT or samples to be measured and anti MT antibody are overlaid and incubate. After a washing step, HRP-conjugated anti rabbit antibody is added and incubate again. HRP conjugated antibody and anti MT antibody MT complex are formed. After a washing, substrate is added, which reacts with the HRP conjugate to produce blue colors. More blue color means less MT. The optical densities are measured and plotted to calculate the exact concentration of MT.
|A. Antigen coated 96well plate||96well plate||1 plate||MT coated plate|
|B. MT standard||1 mL(10μg/mL)||1 vial||Apo Rabbit MT|
|C. Standard diluent||25 mL||1 vial||Phosphatebuffered saline(PBS)|
|D. Antibody solution||12 mL||1 vial||Phosphate buffered saline containing
Anti MT Antibody
|E. 2nd antibody||120 μL||1 vial||HRP-conjughated antibody|
|F. 2nd antibody diluent||12 mL||1 vial||PBS|
|G. Enzyme Substrate||12 mL||1 vial||3,3',5,5'-tetramethylbenz idine(TMB)|
|H. Stop solution||6 mL||1 vial||1N H2SO4|
|I. Concentrated washing solution||50 mL||1 vial||0.2%Tween 20 including x 10 phosphate
|J. Plate seal|
BroadCheck Metallothionein kit