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product information
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| background |
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HSP18.5 is a small heat shock protein localized in the cytoplasm. The protein belongs to HSP20 family. Synonymes:AtHsp18.5, 18.5 kDa heat shock protein |
| immunogen |
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recombinant Arabidopsis thaliana Hsp18.5 |
| antibody format |
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rabbit |
polyclonal |
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serum |
lyophilized |
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| quantity |
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200 µl |
for reconstitution add 200 µl, of sterile water. |
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| storage |
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store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes. |
| tested applications |
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western blot (WB) |
| related products |
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AS08 286 | anti-HSP16.6 | cytosolic class I heat shock protein 16.6, rabbit antibody for cyanobacterial HSP16.6 AS07 254 | anti-HSP17.6 | cytosolic class I heat shock protein 17.6, rabbit antibody
AS08 284 | anti-HSP17.6 | cytosolic class I heat shock protein 17.6, hen antibody AS07 255 | anti-HSP17.7 | cytosolic class II heat shock protein 17.7, rabbit antibody AS08 285 | anti-HSP21 | chloroplastic heat shock protein, rabbit antibody |
| additional information |
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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. |
application information
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| recommended dilution |
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1 : 1000 with standard ECL (WB) |
| expected | apparent MW |
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18.5 kDa |
| confirmed reactivity |
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Arabidopsis thaliana |
| predicted reactivity |
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dicots including: Glycine max, Medicago truncatula, Medicago sativa, Pisum sativum, Ricinus communis, Rosa chinensis, monocots including: Zea mays |
| not reactive in |
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no confirmed exceptions from predicted reactivity are currently known |
| additional information |
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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. |
| selected references |
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to be added when available, antibody released in July 2011. |
application example 
20 µg total protein extracted from 15 day old Arabidopsis thaliana seedlings that were either kept at 22ºC (C) or heat treated at 38ºC for 2 hours in high humidity, prior to protein extraction (HS). As positive control 0.2, 0.4 or 0.8 ng of recombinant Hsp18.5 were separated side by side with the plant samples on 10-15 % gradient SDS-PAGE and
blotted to nitrocellulose (Bio-rad). Blots were blocked following transfer with 5% low fat milk in low salt buffer for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 1000 for 2h at room temperature with agitation in the blocking solution. The primary antibody solution was removed and the blot was rinsed briefly twice, then washed 4 times for 15 min each at room temperature with agitation using low salt buffer. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxides conjugated, from Agrisera, AS09 602), diluted to 1:35 000 for 1h at room temperature with agitation then washed as above and treated with ECL detection reagent according to the manufacturer's instructions.
Exposure time was 20 seconds.
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