cFBPase | Cytosolic fructose-1,6-bisphosphatase (cytoplasm marker in photosynthetic tissues)
AS04 043 | Clonality: polyclonal | Host: Rabbit | Reactivity: A.thaliana, B.napus, M.atropurpureum, N.benthamiana, P.silvestris, O.sativa, P.hybrida cv. Mitchell, S.tuberosum, Z.mays | cellular [compartment marker] of cytoplasm
|Recommended dilution||1 : 5 000 (WB)|
|Expected | apparent MW||
45 | 37 kDa (Arabidopsis thaliana)
|Confirmed reactivity||Arabidopsis thaliana, Brassica napus, Macroptilium atropurpureum, Nicotiana benthamiana, Pinus silvestris, Oryza sativa, Petunia hybrida cv. Mitchell, Solanum tuberosum, Zea mays cellular compartment marker of cytoplasm in photosynthetic tissues
|Predicted reactivity||Capsella rubella, Pisum sativum, Ricinus communis, Glycine max, Phaseolus vulgaris, Sesamum indicum, Spinacia oleracea, Populus trichocarpa, Vitis vinifera|
|Not reactive in||
This antibody does not react with chloroplastic form of FBPase.
|Selected references||Seguel et al. (2018). PROHIBITIN 3 forms complexes with ISOCHORISMATE SYNTHASE 1 to regulate stress-induced salicylic acid biosynthesis in Arabidopsis. Plant Physiol. Jan 2018. DOI:10.1104/pp.17.00941
Lynch et al. (2017). Multifaceted plant responses to circumvent Phe hyperaccumulation by downregulation of flux through the shikimate pathway and by vacuolar Phe sequestration. Plant J. 2017 Dec;92(5):939-950. doi: 10.1111/tpj.13730.
Duan et al. (2017). A Lipid-Anchored NAC Transcription Factor Is Translocated into the Nucleus and Activates Glyoxalase I Expression during Drought Stress. Plant Cell. 2017 Jul;29(7):1748-1772. doi: 10.1105/tpc.17.00044. (Nicotiana benthamiana)
Steffens et al. (2017). Physical, Functional and Genetic Interactions between the BEACH Domain Protein SPIRRIG and LIP5 and SKD1 and Its Role in Endosomal Trafficking to the Vacuole in Arabidopsis. Front Plant Sci. 2017 Nov 20;8:1969. doi: 10.3389/fpls.2017.01969. Xing et al. (2016). Proteome Profile of Starch Granules Purified from Rice (Oryza sativa) Endosperm. PLoS One. 2016 Dec 19;11(12):e0168467. doi: 10.1371/journal.pone.0168467.
LaMontagne et al. (2016). Isolation of Microsomal Membrane Proteins from Arabidopsis thaliana. Curr. Protoc. Plant Biol. 1:217-234. doi: 10.1002/cppb.20020.
Ma et al. (2016). Phosphatidylserine Synthase Controls Cell Elongation Especially in the Uppermost Internode in Rice by Regulation of Exocytosis. PLoS One. 2016 Apr 7;11(4):e0153119. doi: 10.1371/journal.pone.0153119. eCollection 2016.
de Michele et al. (2016). Free-Flow Electrophoresis of Plasma Membrane Vesicles Enriched by Two-Phase Partitioning Enhances the Quality of the Proteome from Arabidopsis Seedlings. J Proteome Res. 2016 Mar 4;15(3):900-13. doi: 10.1021/acs.jproteome.5b00876. Epub 2016 Feb 4.
10 µg of Arabidopsis thaliana Col-0 WT chloroplast total protein (1), 10µg Col-0 WT chloroplast stroma protein (2), Col-0 WT total leaf sample 1:10 dilution (3), Col-0 WT total leaf sample 1:2 dilution (4), Col-0 WT total leaf sample undiluted (5), Col-0 WT total leaf sample undiluted (6) and recombinat plastidial FBPase 0.05 µg, expressed in E.coli with no cTP present in the sequence (7), extracted with 2x Laemmli buffer and denatured at 95°C for 5 min. were separated on 10% SDS-PAGE and blotted to Millipore Immobilon-P membrain (carried out at 100 V for 90 min at 4°C in blotting buffer (25 mM Tris-HCl, 192 mM glycine, 10 % [v/v] methanol). Blots were blocked with blocking solution ( TBST buffer (20 mM Tris-HCl pH 7.4, 150 mM NaCl, 0.05 % [v/v] Tween-20) supplemented with 5 % [w/v] milk powder) for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5 000 for 15h (over noght) at 4°C with agitation in TBST. The antibody solution was decanted and the blot was washed 6 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:20 000 in for 1h at RT with agitation. The blot was washed as above and ChemiGlow West Chemiluminescence substrate (Bucher Biotech, Basel, Switzerland) was used for development according to the manufacturer’s instructions and imaged using the ChemiDoc imaging system (Biorad, Cressier, France). Exposure time was 15 seconds.
Courtesy of Zanella Martina, ETH Zürich, Switzerland
2 µg of total protein from Arabidopsis thaliana roots crude extract (1), supernatant after 10 0000 g centrifugation (2), extracted with ice-cold extraction buffer [50 mM Tris-HCl pH 7.5, 0.33 M Sucrose, 5 mM EDTA, 1x proteinase inhibitor] and denatured gradually with [50 mM Tris (pH 6.8), 10% Glycerol, 2% SDS, 2.5 M Urea, 0.005% Bromophenol Blue] at first with 55°C for 15 min followed by 95°C for 5 min. Proteins were separated on 12 % SDS-PAGE. Proteins were blotted 1h to PVDF using semi-dry transfer. Blots were blocked with 5 % nonfat milk + 0.1 % BSA for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 35 000 for ON/4°C with agitation in TBS-T. 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 RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase) diluted to 1:1 000 in for 2h at RT with agitation. The blot was washed as above and developed for 5 min with SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Thermo Scientific). Exposure time was 20 min.
Courtesy of Dr. Joanna Jeleńska and Dequantarius Speed, University of Chicago, USA
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