Dsp | Death-specific protein
AS10 935 | clonality: polyclonal | host: rabbit | reactivity: diatoms
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1 : 10 000 with standard ECL (WB)
|Expected | apparent MW||
25 | 25 kDa
|Confirmed reactivity||Emiliania huxleyi, Aureococcus anophagefferens, Fragilariopsis cylindrus, Phaeodactylum tricornutum, Skeletonema costatum, Thalassiosira pseudonana
|Not reactive in||
|Selected references||Thamatrakoln et al. (2013). Death-specific protein in a marine diatom regulates photosynthetic responses to iron and light availability. PNAS 2013 Dec 10;110(50):20123-8. doi: 10.1073/pnas.1304727110. Epub 2013 Nov 25.|
5 µg of total protein, extracted with LDS sample buffer (PSB) were separated on 4-20% SDS-PAGE and transferred to PVDF. Blot was blocked with 2% ECL block in TBS-T (0.1% Tween-20) for 1h at room temperature (RT) with agitation. Blot was incubated in primary antibody at a dilution of 1: 10,000 for 1h at RT with agitation. Antibody solution was decanted and the blot rinsed briefly twice with TBS-T, then washed once for 15 min and 3 times for 5 min. at RT with agitation. Blot was incubated in secondary antibody (horse radish peroxidase conjugated anti-rabbit IgG) at a dilution of 1:20 000, for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer’s instructions. Image (20 sec. exposure) captured with a BioRad Chemidoc MP.
Predicted activity and molecular weights (Da) - Thalassiosira pseudonana Dsp1 (24572 Da) Dsp2 (25406 Da), Skeletonema costatum Dsp1 (24848 Da) Dsp2 (26566 Da), Phaeodactylum tricornutum Dsp1 (26585 Da, Fragilariopsis cylindrus Dsp1 (20312 Da).
Competition assay (right half of blot): Same protein loads as for lanes 1-7. Antisera were pre-incubated for 90 min. in TBS-T with of an excess of the free peptide antigen
Courtesy of Dr. K. Thamatrakoln unpublished data; rights retained by the contributor.
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