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

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana BiP proteins: BiP1 At5g28540 Q9LKR3, BiP2 At5g42020 F4K007 , BiP3 At1g09080 Q8H1B3

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum in PBS, pH 7.4

Format Lyophilized

Quantity 50 µg

Reconstitution For reconstitution add 50 µl of sterile water.

Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications ELISA (ELISA), Immunofluorescence (IF), Immunogold (IG), Western blot (WB)

Recommended dilution 1 : 8000 (ELISA), 1 : 600 (IF), 1 : 2000 (WB)

Expected | apparent MW

73.5 | 80 kDa

Reactivity

Confirmed reactivity Anacardium occidentale, Arabidopsis thaliana, Brassica napus, Chara australis R.Br, Chlamydomonas reinhardtii, Cucumis sativus, Mangifera indica, Moniliophthora perniciosa, Nicotiana benthamiana, Nicotiana tabacum, Raphanus sativa L. Tokinashi-daikon, Olea europaea, Oryza sativa, Picea abies, Pistachio sp., Physcomitrella patens, Schinus molle, 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, Petunia hybrida, Picea sitcHensis, Populus trichocarpa, Ricinus comminus, Vitis vinifera
Species of your interest not listed? Contact us

Not reactive in Ostreococcus tauri

Application examples

Application examples Western blot

western blot using plant anti-BiP antibodies

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

Immunofluorescence using plant anti-BiP antibody

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.

Related products

Plant protein extraction buffer

Secondary antibodies

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).