HSP18.5 | class IV heat shock protein
AS11 1628 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Hordeum vulgare
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As hsp18.5 is a low abundancy protein, please, make sure that the plants are heated to the right temperature. Normally I heat stress the seedling on a sealed agar plate for 2 hours. This assures that the humidity around the plant is very high. Low humidity can allow the plant to cool down through transpiration. If a plant is in soil you can keep the leaf or even a whole plant over a wet filter paper and seal the plate very well during the treatment. Heat stressing plants in microphage tubes does not work that well.
Hsp 18.5 is a low abundancy protein and estimated concentration of this protein in total cell extract is ca. 0.007-0.01 %. Therefore to be able to visualize this protein the load per well needs to be at least 20 ug of heat shocked total protein/well. It is crucial to heat treat the plants at 38ºC for 3 hours at high humidity.
Please, note that longer transfer time might result in losing a signal for this protein.
HSP18.5 is a small heat shock protein localized in the cytoplasm. The protein belongs to HSP20 family and is involved in protein complex oligomerization and protein folding. Alternative names: AtHsp18.5, 18.5 kDa heat shock protein.