showForm Anti-BiP | lumenal-binding protein antibodies

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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

BiP | Lumenal-binding protein (rabbit antibody) in the group Antibodies for Plant/Algal  / Membrane Transport System / Endomembrane system at Agrisera AB (Antibodies for research) (AS09 481)

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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, Picea sitcHensis, Populus trichocarpa, Ricinus comminus, Vitis vinifera
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

Related products

AS09 615 | Anti-BiP2 | lumenal-binding protein 2, goat antibodies
AS09 614 | Anti-BiP2 | lumenal-binding protein 2, chicken antibodies
AS09 481PRE | BiP | lumenal-binding protein, pre-immune serum
antibodies to plant endomembrane system proteins

Plant protein extraction buffer

Secondary antibodies

Background

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

Selected references Dalmadi et al. (2019). AGO-unbound cytosolic pool of mature miRNAs in plant cells reveals a novel regulatory step at AGO1 loading. Nucleic Acids Res. 2019 Aug 8. pii: gkz690. doi: 10.1093/nar/gkz690.
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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