BiP | Lumenal-binding protein (100 ĩg)
AS09 481-100| clonality: polyclonal | host: rabbit | reactivity: A. thaliana, B. napus, C. sativus, R. sativa L. Tokinashi-daikon, S. oleracea, S. lycopersicum, S. tuberosum, T. aestivum, Z. mays, O. europaea, P. abies, P. patens, Ch. reinhardtii
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1: 8000 (ELISA), 1: 2000 with standard ECL (WB), 1: 600 (IF)
|Expected | apparent MW||
73.5 | 80 kDa
|Confirmed reactivity||dicots: Arabidopsis thaliana, Brassica napus, Cucumis sativus, Nicotiana benthamiana, Raphanus sativa L. Tokinashi-daikon, Olea europaea, Spinacia oleracea, Solanum lycopersicum, Solanum tuberosum, monocots: Triticum aestivum, Zeay mays, trees: Picea abies, moss: Physcomitrella patens, algae: Chlamydomonas reinhardtii|
Capsella rubella, Vitis vinifera, Ricinus comminus, Glycine max. dicots including: Nicotiana tabacum, monocots: Hordeum vulgare, Oryza sativa, trees: Picea sitchensis, Populus trichocarpa.
|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. This antibody has so far not worked in IP.
|Selected references||Hu et al. (2015). Re-examination of chlorophyllase function implies its involvement in defense against chewing herbivores. Plant Physiol. 2015 Jan 12. pii: pp.114.252023.
Ivanov et al. (2014). SORTING NEXIN1 Is Required for Modulating the Trafficking and Stability of the Arabidopsis IRON-REGULATED TRANSPORTER1. Plant Cell. 2014 Mar 4.
Bommert et al. (2013). The maize Gα gene COMPACT PLANT2 functions in CLAVATA signalling to control shoot meristem size. Nature,Sep 11. doi: 10.1038/nature12583. (western blot, Zea mays)
Sulimanet al. (2013). Cell Polarity and Patterning by PIN Trafficking through Early Endosomal Compartments in Arabidopsis thaliana. PLoS Genet. May;9(5). (immunolocalization).
application example western blot
5 µg of total protein from A.thaliana (1), H. vulgare (2), P. sativum (3)*, Z. mays (4), C. sativus(5), S. tuberosum (6), S. oleracea (7), S. lycopersicum (8) P. patens (9)*, Ch. reinhardtii (10) 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-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) 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. * Lack of the signal or its low signal intensity in those samples can be due to the sample biology. If you work with those species, please inquire.
application example immunolocalization
BiP localization in 5 days old Arabidopsis thaliana roots (A), 3 days old Triticum aestivum roots (B).
BiP signal shown in red, DAPI in blue. The material has been fixed in para-formaldehyde for 30 minutes. Tissue cleaning has been performed before immunolocalization. Rabbit anti-BiP primary antibody diluted in 1: 600 and ALEXA 555 conjugated anti-rabbit secondary antibody (red color) have been used. Co-staining with DAPI visualized nucleus (blue color). Scale bar – 10 µm.
Courtesy Dr. Taras Pasternak, Freiburg University
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