HSP70 | Heat shock protein 70 (mitochondrial)

Product no: AS08 347

AS08 347 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thalian, Brassica oleracea, Hordeum vulagre

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  • Product Info
  • Immunogen:

    KLH-conjugated peptide conserved in higher plant mitochondrial HSC70 including Arabidopsis thaliana mtHSC70-1 Q8GUM2, At4g37910 and mtHSC70-2 Q9LDZ0, At5g09590

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Immunogen affinity purified serum in PBS pH 7.4.
    Format: Lyophilized
    Quantity: 2 x 50 g
    Reconstitution: For reconstitution add 50 µl of sterile water per tube
    Storage: Store lyophilized/reconstituted at -20C; 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: Western blot (WB)
    Recommended dilution: 1 : 1000 (WB)
    Expected | apparent MW:

    73 | 70 kDa

  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Brassica oleracea, Hordeum vulgare
    Predicted reactivity: Medicago truncatula, Oryza sativa, Phaseolus vulgaris, Pisum sativum, Populus trichocarpa, Spinacia oleracea, Solanum tuberosum, Triticum aestivum, Zea mays, Vitis vinifera
    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Application example

    western blot using anti-HSP70 mitochondrial antibodies

    25 μg of Arabidopsis thaliana leaf extract and 15 μg)Arabidopsis thaliana mitochondrial fraction were separated on 10% gel and blotted on nitrocellulose membrane using wet transfer (0.22% CAPS, pH 11). Filters where blocked (1.5h) in 5% milk in TBST (1X TBS, 0,1% Tween 20), incubated with 1: 1000 anti-HSP70 mitochondrial antibodies (2h in TBST) followed by incubation with 1: 10 000 secondary anti-rabbit (1h) HRP-coupled antibodies and visualized with standard ECL on Kodak autoradiography film for 15-60 s. Mitochondria were isolated as described by Urantowka et al. (Plant Mol Biol, 2005, 59:239-52). Mitochondrial pellets were suspended in 1X Laemmli buffer (5% beta-mercaptoetanol, 3.7% glycerol, 1.1% SDS, 23 mM Tris- HCl pH 6.8, 0.01% bromophenol blue), heated (95°C, 5 min.) and centrifuged (13 000rpm, 1 min.). Leaf extracts were prepared as described by Martinez-Garcia et al. (Plant J., 1999, 20:251-7).

    Courtesy Dr. Janusz Piechota, Wrocław University, Poland


  • Background
  • Background:

    Heat-shock protein 70 (Hsp70) is the major stress-inducible protein in vertebrates and is highly conserved throughout evolution. It plays a role as a molecular chaperone and is important for allowing cells to cope with acute stressor insult, especially those affecting the protein machinery. Heat shock cognate protein 70 (HSC70), is a highly conserved protein and a member of the family of molecular chaperones.

  • Product Citations
  • Selected references: Lentini et al. (2018). Early responses to cadmium exposure in barley plants: effects on biometric and physiological parameters. Acta Physiol Plant (2018) 40: 178. https://doi.org/10.1007/s11738-018-2752-2.
    Rurek et al. (2018). Mitochondrial Biogenesis in Diverse Cauliflower Cultivars under Mild and Severe Drought Involves Impaired Coordination of Transcriptomic and Proteomic Response and Regulation of Various Multifunctional Proteins. Preprints 2018, 2018010276 (doi: 10.20944/preprints201801.0276.v1).
    Opalińska et al. (2017). Identification of Physiological Substrates and Binding Partners of the Plant Mitochondrial Protease FTSH4 by the Trapping Approach. Int J Mol Sci. 2017 Nov 18;18(11). pii: E2455. doi: 10.3390/ijms18112455.
    Murcha et al. (2016). Plant specific Preprotein and Amino Acid Transporter proteins are required for tRNA import into mitochondria. Plant Physiol. 2016 Oct 27. pii: pp.01519.2016.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts
  • Reviews:
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    Michal RUREK | 2014-02-14
    0 | 0
    20 g of mitochondrial proteins from cauliflower (Brassica oleracea var. botrytis) curds (apical layer) was separated on 12 % SDS-PAGE (Laemmli- type) and blotted 1h to Immobilone P (Millipore) using Sedryt apparatus (Kucharczyk). After blocking (5% milk in PBS-T) for 1 h at RT, blots were incubated in the primary antibody at a dilution of 1: 4000 for overnight at 4 deg. with agitation. The primary antibody, diluted in 2% milk in PBS-T was reused several times. Secondary, anti-rabbit HRP- linked antibodies were bound at 1/10000 dilution at RT (1h). Single band of ca. 70 kDa was detected using standart GE Healthcare ECL reagents.

Accessories

AS08 371 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, B. juncea, B. oleracea, C. sativus, C. reinhardtii, D. subspicatus, E. tef, G. vermiculophylla, H. vulgare, M. sativa, O. sativa, P. kurroa, P. strobus, Salicornia sp., S. italica. S. vulgaris, S. lycopersicum, Trebouxia TR1 and TR9, T. aestivum, Z. mays, P. falciparum, V. faba

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302 €
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AS16 ECL-S-N | low pico to mid femtogram and extreme low femtogram detection

Now available only as separate products:
1 x AS16 ECL-N
1 x AS16 ECL-S

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