BiP | Lumenal-binding protein (rabbit antibody)
AS09 481| clonality: polyclonal | host: rabbit | reactivity: A.thaliana, B.napus, C.reinhardtii, C.sativus, M.perniciosa, N.benthamiana, N.tabacum, R.sativa L. Tokinashi-daikon, O.europaea, O.sativa, P.abies, P.patens, S.oleracea, S. lycopersicum, S.tuberosum, T.aestivum, Z.mays
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|Recommended dilution||1 : 8000 (ELISA), 1 : 600 (IF), 1 : 2000 (WB)|
|Expected | apparent MW||
73.5 | 80 kDa
|Confirmed reactivity||Arabidopsis thaliana, Brassica napus, Chlamydomonas reinhardtii, Cucumis sativus, Moniliophthora perniciosa, Nicotiana benthamiana, Nicotiana tabacum, Raphanus sativa L. Tokinashi-daikon, Olea europaea, Oryza sativa, Picea abies, Physcomitrella patens, Spinacia oleracea, Solanum lycopersicum, Solanum tuberosum, Triticum aestivum, Zea mays|
|Predicted reactivity||Arabis alpina, Capsella rubella, Capsicum annuum, Citrus clementina, Citrus sinsensis, Eucalyptus grandis, Glycine max, Hordeum vulgare, Isatis tincorina, Prunus persica, Triticum aestivium, Picea sitcHensis, Populus trichocarpa, Ricinus comminus, Vitis vinifera|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
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||Mares et al. (2017). Proteomic analysis during of spore germination of Moniliophthora perniciosa, the causal agent of witches' broom disease in cacao. BMC Microbiol. 2017 Aug 17;17(1):176. doi: 10.1186/s12866-017-1085-4.
Gelová et al. (2017). Antibody-mediated modulation of cytokinins in tobacco: organ-specific changes in cytokinin homeostasis. J Exp Bot. 2017 Dec 23. doi: 10.1093/jxb/erx426.
Nagel et al. (2017). Arabidopsis SH3P2 is an ubiquitin-binding protein that functions together with ESCRT-I and the deubiquitylating enzyme AMSH3. Proc Natl Acad Sci U S A. 2017 Aug 7. pii: 201710866. doi: 10.1073/pnas.1710866114.
Lomin et al. (2017). Studies of cytokinin receptor–phosphotransmitter interaction provide evidences for the initiation of cytokinin signalling in the endoplasmic reticulum. Functional Plant Biology, CSIRO Publications. (Nicotiana benthamiana, western blot)
Zhang et al. (2017). Control of secondary cell wall patterning involves xylan deacetylation by a GDSL esterase. Nat Plants. 2017 Mar 3;3:17017. doi: 10.1038/nplants.2017.17. (Oryza sativa, immunolocalization, western blot)
Je et al. (2016). Signaling from maize organ primordia via FASCIATED EAR3 regulates stem cell proliferation and yield traits. Nat Genet. 2016 Jul;48(7):785-91. doi: 10.1038/ng.3567. Epub 2016 May 16.
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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