BiP | Lumenal-binding protein (rabbit antibody)

Product no: AS09 481

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 Info
  • 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 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 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, Schizosaccharomyces pombe
  • 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

    Application examples:

    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 (±SD) 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
  • Additional information (application): 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
  • 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: Miloro et al. (2024). Barley AGO4 proteins show overlapping functionality with distinct small RNA-binding properties in heterologous complementation. Plant Cell Rep. 2024 Mar 13;43(4):96. doi: 10.1007/s00299-024-03177-z.  
    Xue et al.(2023). The PtdIns3P phosphatase MtMP promotes symbiotic nitrogen fixation via mitophagy in Medicago truncatula. iScience. 2023 Sep 15;26(10):107752.doi: 10.1016/j.isci.2023.107752.
    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.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Posters Collection



    Method of isolation of plant ER

    methd of plant ER isolation

    method of isolation of plant ER (2)

     

    Courtesy of Dr. Masayoshi Maeshima, Laboratory of Cell Dynamics, Graduate School of Bioagricultural Sciences Nagoya University Nagoya, Japan

  • Reviews:
  • Reviews
    Below, you can grade the product on a scale from 0 to 5.
    Please also provide information about species, application, dilution and obtained result for the reviewed antibody.
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    Number of reviews: (4)
    Alberto Dvalos | 2024-04-12
    0 | 0
    This antibody worked very well for A. thaliana Extracellular Vesicles and from Brassica oleracea (broccoli) EVs. Dilution 1:2000 in 10% SDS PAGE
    Peter Ma | 2021-02-04
    0 | 0
    This antibody works very well for our samples. The membrane proteins from the Arabidopsis roots are subjected to 10% SDS-PAGE and then detected with this antibody (1:2000). The results shows that this antibody is quite specific and sensitive.
    Francesca | 2014-11-09
    0 | 0
    We successfully used Bip antibody diluted 1:10000 against S. lycopersicon leaves extracts. Probing was conducted for 1h in PBS 5% skim milk.
    Sidona | 2011-11-16
    0 | 0
    We have used BiP2 antibody with protein extract from Nicotiana and Malus species at dilution 1:2000 . The same dilution could be re-used 2 times.

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