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



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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 Immunogen affinity purified serum in PBS pH 7.4.
Format Lyophilized
Quantity 50 ĩg
Reconstitution For reconstitution add 50 ĩl of sterile water
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
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., Physcomitrium 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

Reactant: Malus domestica (Apple)

Application: Western Blotting

Pudmed ID: 23886449

Journal: Plant Methods

Figure Number: 2A

Published Date: 2013-07-28

First Author: Sikorskaite, S., Rajamäki, M. L., et al.

Impact Factor: 5.139

Open Publication

Analysis of protein fractions from different steps of nuclei isolation. The protein fractions obtained following the main steps of the procedure used for isolation of nuclei from leaves of apple are shown. Equal amounts of proteins (4 ?g) from each fraction were loaded on the gel. A) Lumenal-binding protein 2 (BiP2), B) histone H3 and C) plastocyanin (PC) were detected with specific antibodies by western blot analysis. D) Coomassie blue staining of the fractionated proteins separated by polyacrylamide gel electrophoresis. Lane 1: crude extract of proteins from homogenized leaf tissue; lane 2: resuspended pellet of the whole cell lysate obtained following treatment with Triton X-100 and centrifugation; lane 3: nuclei collected from the 60% Percoll layer (see nuclear fraction in Figure 1B); lane 4: the interface fraction of 60% Percoll and 2.5 M sucrose layers in the density gradient containing chloroplasts and unbroken cells (see 60% P/2.5 M S interface in Figure 1B); lane 5: sucrose layer of the density gradient (see 2.5 M sucrose layer in Figure 1B); lane 6: nuclei purified by centrifugation on a 35% Percoll cushion.


Reactant: Solanum lycopersicum (Tomato)

Application: Western Blotting

Pudmed ID: 26347766

Journal: Front Plant Sci

Figure Number: 6A

Published Date: 2015-09-09

First Author: Buxa, S. V., Degola, F., et al.

Impact Factor: 5.435

Open Publication

(A–D) Western blot analyses of S. lycopersicum midrib extracts. Three healthy (lanes 1–3) and three infected (lanes 4–6) plants were analyzed. Immunoreactive signals corresponding to actin and ER luminal binding protein (BiP) displayed the expected molecular weights of ?40 and ?78 kDa, respectively (A). Amount of loaded protein were checked by Ponceau-S staining (B). Quantification of immunoreactive luminescence signals was achieved by densitometric scanning of the respective actin (C) and BiP (D) bands. The mean of six replicates (ąSD) was calculated for each sample. Actin T-value: –9.977; BiP T-value: 15.068. * denotes significant difference of the means (P-values ? 0.001).


Reactant: Moniliophthora perniciosa (Witches' broom disease)

Application: Western Blotting

Pudmed ID: 28818052

Journal: BMC Microbiol

Figure Number: 4A

Published Date: 2017-08-17

First Author: Mares, J. H., Gramacho, K. P., et al.

Impact Factor: 3.437

Open Publication

Accumulation of BiP, Catalase, and ATP synthase (beta chain) by western blot and analysis of spots. a Image of a nitrocellulose membrane hybridized with Anti-BiP, Anti-Catalase, and Anti-ATP synthase (beta chain) of Arabidopsis (Normalization, see Additional file 6: Figure S4). b Relative accumulation determined from images of the nitrocellulose membrane, using the Gel Quant. Net 1.8.0 software. c Relative quantification of each spot corresponding to proteins analyzed by western blot. The average values of the each spot’s percentage volume in each gel replicate was used for graph construction


Reactant: Chlamydomonas reinhardtii (Green Alga)

Application: Western Blotting

Pudmed ID: 32731621

Journal: Cells

Figure Number: 6A

Published Date: 2020-07-28

First Author: Garrido, C., Caspari, O. D., et al.

Impact Factor: None

Open Publication

Biochemical confirmation of AMP targeting activity. Mitochondrial, whole cell and chloroplast fractions (1 ?g protein per well) isolated from Chlamydomonas strains in which Venus localization is driven by bacillocin 1580 (A) or magainin II (B), each fused to the RBCA cleavage site, were immunolabelled with antibodies raised against FLAG, an epitope tag carried C-terminally by the Venus reporter, and markers for different cellular compartments: Cytochrome Oxidase subunit IIb (COXIIb) and ATPsynthase subunit F1? for mitochondria (mt), Photosystem II Oxygen Evolving Enhancer 2 (OEE2) and Rubisco small subunit (RBCS) for chloroplasts (cp), ?-Tubulin and nucleic-acid binding protein 1 (NAB1) for the cytosol (cyt) and luminal binding protein (BiP) for the endoplasmic reticulum (ER). Isolated chloroplasts from the Bacillocin 1580 strain (C) and isolated mitochondria from the Magainin II strain (D) were subjected to a proteinase assay, where aliquots were treated with either 150 ?g mL?1 proteinase K and/or 1% Triton X-100, a membrane solubilizing detergent. Aliquots were subsequently immuno-labelled with antibodies against FLAG, chloroplast ATP synthase subunit CF1 ? and other organelle-markers described aside.


Reactant: Chlamydomonas reinhardtii (Green Alga)

Application: Western Blotting

Pudmed ID: 32731621

Journal: Cells

Figure Number: 6C

Published Date: 2020-07-28

First Author: Garrido, C., Caspari, O. D., et al.

Impact Factor: None

Open Publication

Biochemical confirmation of AMP targeting activity. Mitochondrial, whole cell and chloroplast fractions (1 ?g protein per well) isolated from Chlamydomonas strains in which Venus localization is driven by bacillocin 1580 (A) or magainin II (B), each fused to the RBCA cleavage site, were immunolabelled with antibodies raised against FLAG, an epitope tag carried C-terminally by the Venus reporter, and markers for different cellular compartments: Cytochrome Oxidase subunit IIb (COXIIb) and ATPsynthase subunit F1? for mitochondria (mt), Photosystem II Oxygen Evolving Enhancer 2 (OEE2) and Rubisco small subunit (RBCS) for chloroplasts (cp), ?-Tubulin and nucleic-acid binding protein 1 (NAB1) for the cytosol (cyt) and luminal binding protein (BiP) for the endoplasmic reticulum (ER). Isolated chloroplasts from the Bacillocin 1580 strain (C) and isolated mitochondria from the Magainin II strain (D) were subjected to a proteinase assay, where aliquots were treated with either 150 ?g mL?1 proteinase K and/or 1% Triton X-100, a membrane solubilizing detergent. Aliquots were subsequently immuno-labelled with antibodies against FLAG, chloroplast ATP synthase subunit CF1 ? and other organelle-markers described aside.

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

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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 Gu et al. (2021) A Lipid Bodies-Associated Galactosyl Hydrolase Is Involved in Triacylglycerol Biosynthesis and Galactolipid Turnover in the Unicellular Green Alga Chlamydomonas reinhardtii
Dittmer, Kleine, & Schwenkert. (2021) The TPR- and J-domain-containing proteins DJC31 and DJC62 are involved in abiotic stress responses in Arabidopsis thaliana. J Cell Sci. 2021 Oct 1;134(19):jcs259032. doi: 10.1242/jcs.259032. Epub 2021 Oct 12. PMID: 34515300.
Shteinberg et al. (2021) Tomato Yellow Leaf Curl Virus (TYLCV) Promotes Plant Tolerance to Drought. Cells. 2021 Oct 25;10(11):2875. doi: 10.3390/cells10112875. PMID: 34831098; PMCID: PMC8616339.
Mishra et al. (2021) Interplay between abiotic (drought) and biotic (virus) stresses in tomato plants. Mol Plant Pathol. 2021 Dec 30. doi: 10.1111/mpp.13172. Epub ahead of print. PMID: 34970822.
Yang et al. (2020). PROTEIN PHOSPHATASE 95 Regulates Phosphate Homeostasis by Affecting Phosphate Transporter Trafficking in Rice. Plant Cell. 2020 Jan 9. pii: tpc.00685.2019. doi: 10.1105/tpc.19.00685.
Jang et al. (2020). 1Molecules and CellsCrABCA2 Facilitates Triacylglycerol Accumulation in Chlamydomonas reinhardtii under Nitrogen Starvation. Mol Cells. 2020 Jan 31;43(1):48-57. doi: 10.14348/molcells.2019.0262.
Hurny et al. (2020). SYNERGISTIC ON AUXIN AND CYTOKININ 1 Positively Regulates Growth and Attenuates Soil Pathogen Resistance. Nat Commun. 2020 May 1;11(1):2170. doi: 10.1038/s41467-020-15895-5. (immunolocalization)
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.
Mares et al. (2020). Hydrosoluble phylloplane components of Theobroma cacao modulate the metabolism of Moniliophthora perniciosa spores during germination.Fungal Biol. 2020 Jan;124(1):73-81. doi: 10.1016/j.funbio.2019.11.008.
Zhang et al. (2019) Arabinosyl Deacetylase Modulates the Arabinoxylan Acetylation Profile and Secondary Wall Formation. Plant Cell. 2019 May;31(5):1113-1126. doi: 10.1105/tpc.18.00894. Epub 2019 Mar 18. PMID: 30886126; PMCID: PMC6533017.
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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