About: Using of chicken antibodies for metallothionein detection in human blood serum and cadmium-treated tumour cell lines after dot- and electroblotting     Goto   Sponge   NotDistinct   Permalink

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  • Metallothionein (MT) is a protein playing role in homeostasis of metal ions. Its relation with formation and progression of tumors is discussed. Here, we propose a new methodological approach for visualization of MT on PVDF membranes after dot- and electroblotting by using a commercial monoclonal antibody and polyclonalchicken antibodies. The optimized procedure was as follows. We dotted sample on membrane and let it to dry. Then, we blocked the membrane with BSA. After that, the membrane was incubated in chicken primary antibody, washed, and incubated in secondary antibody conjugated with horseradish peroxidase. To visualize, we used 3-aminoethyl-9-carbazole. Under these conditions, we estimated detection limit as 3 pg of MT per 1 microL. The optimal approach was further utilized for detection of MT level in 2 human fibroblast lines and in serum obtained from children with medulloblastoma. The results were in good agreement with differential pulse voltammetry –Brdicka reaction.
  • Metallothionein (MT) is a protein playing role in homeostasis of metal ions. Its relation with formation and progression of tumors is discussed. Here, we propose a new methodological approach for visualization of MT on PVDF membranes after dot- and electroblotting by using a commercial monoclonal antibody and polyclonalchicken antibodies. The optimized procedure was as follows. We dotted sample on membrane and let it to dry. Then, we blocked the membrane with BSA. After that, the membrane was incubated in chicken primary antibody, washed, and incubated in secondary antibody conjugated with horseradish peroxidase. To visualize, we used 3-aminoethyl-9-carbazole. Under these conditions, we estimated detection limit as 3 pg of MT per 1 microL. The optimal approach was further utilized for detection of MT level in 2 human fibroblast lines and in serum obtained from children with medulloblastoma. The results were in good agreement with differential pulse voltammetry –Brdicka reaction. (en)
Title
  • Using of chicken antibodies for metallothionein detection in human blood serum and cadmium-treated tumour cell lines after dot- and electroblotting
  • Using of chicken antibodies for metallothionein detection in human blood serum and cadmium-treated tumour cell lines after dot- and electroblotting (en)
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  • Using of chicken antibodies for metallothionein detection in human blood serum and cadmium-treated tumour cell lines after dot- and electroblotting
  • Using of chicken antibodies for metallothionein detection in human blood serum and cadmium-treated tumour cell lines after dot- and electroblotting (en)
skos:notation
  • RIV/00064203:_____/09:5392!RIV10-MZ0-00064203
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • V
http://linked.open...iv/cisloPeriodika
  • 21
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
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  • 348001
http://linked.open...ai/riv/idVysledku
  • RIV/00064203:_____/09:5392
http://linked.open...riv/jazykVysledku
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  • Cancer cell lines; Differential pulse voltammetry-Brdicka reaction; Dot immunobinding assay; Metallothionein; Western blot; performance liquid-chromatography; transfer stripping technique; immunosorbent-assay elisa; messenger-rna expression; mercury saturation assay; capillary-electrophoresis; brdicka reaction; electrochemical detection; mass-spectrometry; eisenia-foetida (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • DE - Spolková republika Německo
http://linked.open...ontrolniKodProRIV
  • [190E57A5EDEA]
http://linked.open...i/riv/nazevZdroje
  • Electrophoresis
http://linked.open...in/vavai/riv/obor
http://linked.open...ichTvurcuVysledku
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http://linked.open...UplatneniVysledku
http://linked.open...v/svazekPeriodika
  • 30
http://linked.open...iv/tvurceVysledku
  • Adam, V.
  • Eckschlager, Tomáš
  • Kizek, R.
  • Krizkova, S.
http://linked.open...ain/vavai/riv/wos
  • 000272126000011
issn
  • 0173-0835
number of pages
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