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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73.5 | 80 kDa
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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.
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
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.
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).
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