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BiP | Lumenal-binding protein (rabbit antibody)

272 €
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AS09 481| clonality: polyclonal | host: rabbit | reactivity: A.thaliana, B.napus, 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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AS09 481

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product information
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).

Immunogen

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

Host Rabbit
Clonality Polyclonal
Clone
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)
Related products

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

Plant protein extraction buffer

Secondary antibodies

Additional information

Method for plant ER isolation is available here.

application information
Recommended dilution 1 : 8000 (ELISA), 1 : 600 (IF), 1 : 2000 (WB)
Expected | apparent MW

73.5 | 80 kDa

Confirmed reactivity Arabidopsis thaliana, Brassica napus, Chlamydomonas reinhardtii, Cucumis sativus, Moniliophthora perniciosa, Nicotiana benthamiana, Nicotiana tabacum, Raphanus sativa L. Tokinashi-daikon, Olea europaea, Oryza sativa, Picea abies, Physcomitrella patens, 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 No confirmed exceptions from predicted reactivity are currently known.
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.

Selected references 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.

Application example 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)*Ch. 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  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 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.

Application example 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


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