HSP70 | Heat shock protein 70 cytoplasmic
AS08 371 | clonality:polyclonal | host:rabbit | reactivity:A. thaliana, C. sativus, C. reinhardtii, D. subspicatus, E. tef, G. vermiculophylla, H. vulgare, M. sativa, P. strobus, S. vulgaris, S. lycopersicum, Trebouxia TR1 and TR9, Z. mays, P. falciparum
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1: 3000 with standard ECL on 5 µg of protein per well (WB), 2-3 µl/protein extract of concentration 3-5 mg/ml
|Expected | apparent MW||
Arabidopsis thaliana, Chlamydomonas reinhardtii, Chlamydomonas sp. UWO241, Cucumis sativus, E. teft, Hordeum vulgare, Medicago sativa, Silene vulgaris, Solanum lycopersicum, Pinus strobus, Polyscias elegans, Zea mays, algae: Desmodesmus subspicatus, Gracilaria vermiculophylla, phycobiont: Trebouxia TR1 and TR9, Plasmodium falciparum
Ageratina adenophora, Allium sativum, Arabis alpina, Arachis diogoi, Arundo donax, Brassica napus, brassica rapa subsp. pekinensis, Camellia sinensis, Citrus sinensis, Coffea arabica, Eriobotrya japonica, Gossypium arboretum, Glycine max, Glycine soja, Hordeum vulgare var. distichum, Lotus japonicus, Medicago sativa, medicago truncatula, Musa acuminata subsp. malaccensis, Nicotiana tabacum, Nicotiana bethamiana, Oryza sativa, Phaseolus vulgaris, Physcomitrella patens, Pinus taeda, Pisum sativum, Populus balsamifera, Populus trichocarpa, Salix gilgiana, Saussurea medusa,Solanum tuberosum, Solanum commersonii, Spinacia oleracea, Tragopogon dubius, Tragopogon porrifolius, Triticum aestivum, Vitis vinifera, Volvox sp.
|Not reactive in||
No confirmed exceptions from predicted reactivity known in the moment
This product can be sold containing ProClin if requested
|Selected references||Gorovits et al. (2017). The six Tomato yellow leaf curl virus genes expressed individually in tomato induce different levels of plant stress response attenuation. Cell Stress Chaperones. 2017 Mar 21. doi: 10.1007/s12192-017-0766-0.
Fernández-Bautista N. et al. (2017). AtHOP3, a member of the HOP family in Arabidopsis, interacts with BiP and plays a major role in the ER stress response. Plant Cell Environ. 2017 Feb 2. doi: 10.1111/pce.12927.
Hammann et al. (2016). Selection of heat‑shock resistance traits during the invasion of the seaweed Gracilaria vermiculophylla. Marine Biology 163: 104.
Shen et al. (2016). The Arabidopsis polyamine transporter LHR1/PUT3 modulates heat responsive gene expression by enhancing mRNA stability. Plant J. 2016 Aug 19. doi: 10.1111/tpj.13310. [Epub ahead of print]
Gorovits et al. (2016). Tomato yellow leaf curl virus confronts host degradation by sheltering in small/midsized protein aggregates. Virus Res. 2016 Feb 2;213:304-13. doi: 10.1016/j.virusres.2015.11.020. Epub 2015 Dec 1.
Ghandi et al. (2016). Tomato yellow leaf curl virus infection mitigates the heat stress response of plants grown at high temperature. Sci Rep. 2016 Jan 21;6:19715. doi: 10.1038/srep19715.
Kim et al. (2015). Cytosolic targeting factor AKR2A captures chloroplast outer membrane-localized client proteins at the ribosome during translation. Nat Commun. 2015 Apr 16;6:6843. doi: 10.1038/ncomms7843.
Hattab et al. (2015). Characterisation of lead-induced stress molecular biomarkers in Medicago sativa plants. Environm. Exp. Botany. Volume 123, March 2016, Pages 1–12.
Derbyshire et al. (2015). Proteomic Analysis of Microtubule Interacting Proteins over the Course of Xylem Tracheary Element Formation in Arabidopsis. Plant Cell. 2015 Oct 2. pii: tpc.15.00314.
Law et al. (2015). Phosphorylation and Dephosphorylation of the Presequence of pMORF3 During Import into Mitochondria from Arabidopsis thaliana. Plant Physiol. 2015 Aug 24. pii: pp.01115.2015.
Moshe at al. (2015). Tomato plant cell death induced by inhibition of HSP90 is alleviated by Tomato yellow leaf curl virus infection. Mol Plant Pathol. 2015 May 12. doi: 10.1111/mpp.12275.
Piechota et al. (2015). Unraveling the functions of type II-prohibitins in Arabidopsis mitochondria. Plant Mol Biol. 2015 Apr 21.
Hazlina et al. (2015). Photoinhibition and Development of Stress Proteins in Macroalgae Irradiated with Ultraviolet Radiation. ASM Sci. J., 7(2), 118–128.
Guggisberg et al. (2014). A sugar phosphatase regulates the methylerythritol phosphate (MEP) pathway in malaria parasites. Nat Commun. 2014 Jul 24;5:4467. doi: 10.1038/ncomms5467.
Liu et al. (2014). Spermidine Enhances Waterlogging Tolerance via Regulation of Antioxidant Defence, Heat Shock Protein Expression and Plasma Membrane H+-ATPase Activity in Zea mays. J. Agronomy and Crop Science, Article first published online: 1 APR 2014, DOI: 10.1111/jac.12058.
1µg of total protein from (1) Horderum vulgare pre heat shock leaf extracted with PEB (AS08 300), (2) Horderum vulgare post heat shock (2h 40ºC) leaf extracted with PEB (AS08 300), (3) Zea mays pre heat shock total protein leaf extracted with PEB (AS08 300), (4) Zea mays post heat shock (2h 40ºC) total protein leaf extracted with PEB (AS08 300) were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF (Milipore). Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-HSP70 antibody (AS08 371, 1:20 000, 1h) and secondary anti-rabbit (1:20 000, 1 h) antibody (HRP conjugated) in TBS-T containing 2% low fat milk powder. All steps were performed at RT with agitation. Signal was detected withECL Advance (GE Healthcare)
Protein from Solanum lycopersicum (1) total cell extract ca. 30 -50 µg, (2) and (3) nuclei pellet , (4) and (5) ca. 7 µg of nuclei fraction, (6) and (7) cytoplasmic pellet, (8) ca. 7 µg of cytoplasm fraction, were separated on 10% SDS-PAGE and blotted 1h to nitrocellulose (Schleicher & Schuell). Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-HSP70 antibody (AS08 371, 1:5000, 3h RT). The antibody solution was decanted and the blot was rinsed briefly. Washed 3 times for 15 min in TBS-T at room temperature with agitation. Blot was incubated with a secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 5:000. The blot was washed as above and developed for 1 min with ECL detection reagent according to the manufacturers instructions.
Courtesy Dr Rena Gorovits
|200 fmoles of HSP70 protein standard (1), product number AS08 371S, 1 µg of total protein from samples such as Lycopersicum esculentum leaf (2), Nicotiana tabaccum leaf, (3), Zea mays leaf (4), Hordeum vulgare leaf (5), Arabidopsis thaliana leaf (6) were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70°C for 5 min and keept on ice before loading. Protein samples were separated on 4- 12% Bolt Plus gels, LDS-PAGE and blotted for 70 minutes to PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent (GE RPN 2125; Healthcare) or 5% non-fat milk dissolved in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 10 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS10 1489, Agrisera) diluted to 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with TMA-6 (Lumigen) detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (VersaDoc MP 4000) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.||
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