BiP | lumenal-binding protein
AS09 615 | clonality: polyclonal | host: goat | reactivity: Arabidopsis thaliana, Spinacia oleracea
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1: 2000 with standard ECL (WB)
|Expected | apparent MW||
73.5 | 80 kDa
|Confirmed reactivity||dicots including: Arabidopsis thaliana, Spinacia oleracea, monocots including: Hordeum vulgare, Zea mays
dicots including: Nicotiana tabacum, Spinacia oleracea, monocots: Hordeum vulgare, Oryza sativa, Zea mays, trees: Picea sitchensis, Populus trichocarpa, moss: Physcomitrella patens
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.
Antibody has a reduced reactivity to monocots in western blot.
|Selected references||Narusaka et al (2016). Leucine zipper motif in RRS1 is crucial for the regulation of Arabidopsis dual resistance protein complex RPS4/RRS1. Sci Rep. 2016 Jan 11;6:18702. doi: 10.1038/srep18702.|
5 µg of total protein from A.thaliana (1), H. vulgare (2), Z.mays (3), S. oleracea (4), extracted with Agrisera PEB extraction buffer (AS08 300) were separated on 4-12% SDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 10 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-goat IgG horse radish peroxidase conjugated, from Agrisera AS09 605) diluted to 1:50 000 for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL detection reagent according to the manufacturers instructions. Exposure time was 5 seconds.
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