BiP | Lumenal-binding protein (rabbit antibody)
AS09 481 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A.thaliana, B.napus, Chara australis 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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Product Information
KLH-conjugated synthetic peptide derived from Arabidopsis thaliana BiP proteins: BiP1 At5g28540 Q9LKR3, BiP2 At5g42020 F4K007 , BiP3 At1g09080 Q8H1B3
73.5 | 80 kDa
Reactivity
Application examples
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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)*, C. 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 5 % non-fat milk in TBS-T, 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 of extreme femtogram range, 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.
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, Germany
Additional information
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.
Background
BiP2 (Binding immunoglobulin protein) is localized in endoplasmic reticulum lumen (ER) and plays a role in protein assembly inside ER. BiP protein is abundant under all growth conditions but its synthesis can increase under conditions that lead to the accumulation of unfolded polypeptides in endoplasmic reticulum (ER).
Product citations
Feng et al. (2019). Analyses of transgenic fibroblast growth factor 21 mature rice seeds. J-STAGe, Online ISSN : 1347-3735.
Bastiaan-Net et al. (2018). IgE Cross-Reactivity of Cashew Nut Allergens. Int Arch Allergy Immunol. 2018 Oct 26:1-14. doi: 10.1159/000493100.
Wang et al. (2018). Resistance protein Pit interacts with the GEF OsSPK1 to activate OsRac1 and trigger rice immunity. Proc Natl Acad Sci U S A. 2018 Nov 16. pii: 201813058. doi: 10.1073/pnas.1813058115.
Pertl-Obermeyer et al. (2018). Dissecting the subcellular membrane proteome reveals enrichment of H+ (co-)transporters and vesicle trafficking proteins in acidic zones of Chara internodal cells. PLoS One. 2018 Aug 29;13(8):e0201480. doi: 10.1371/journal.pone.0201480.
Qiao et al. (2018). Two Crinivirus-Conserved Small Proteins, P5 and P9, Are Indispensable for Efficient Lettuce infectious yellows virus Infectivity in Plants. Viruses. 2018 Aug 28;10(9). pii: E459. doi: 10.3390/v10090459.
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.
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