FtsH10 | ATP-dependent zinc metalloprotease FtsH10 (mitochondrial)

Product no: AS07 251

AS07 251 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

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  • Product Info
  • Immunogen:

    KLH-conjugated peptide located near C-terminus chosen from sequence of Arabidopsis thaliana FtsH10 Q8VZI8, At1g07510

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Serum
    Format: Lyophilized
    Quantity: 200 l
    Reconstitution: For reconstitution add 200 l of sterile water
    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: Blue Native PAGE (BN-PAGE), Immunoprecipitation (IP)
    Recommended dilution: 1 : 1000 (WB)
    Expected | apparent MW:

    84 kDa

  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Predicted reactivity:

    Arabidopsis thaliana

    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Application example


    western blot detection using anti-FtsH10 antibodies

     

    Mitochondrial preapration from Arabidopsis thaliana mitochondria was separated on 10% acrilamide gel and electrophoresis prepared according to Schägger and von Jagov (Anl. Biochem., 1987, 166:368-379). After running the gel, proteins were transferred to nitrocellulose membrane using wet transfer (0.22% CAPS, pH 11). Transfer was checked by Ponceau S staining. Blot was destained by several quick washings in distilled water and 1 washing in 1X TBS (10 mM T pH 7.5, 150 mM NaCl) (10-15 min.).Blot was blocked by 1.5 hour in 5% milk in TBST (1X TBS, 0,1 20) After blocking blot was washed quickly twice in TBST and incubated 2 hours with primary antibody (dilution 1: 1000 TBST (dilution 1:1000). Washing: two quick washings in TBST and 3 x 10 min. washings in TBST. Then blot was incubated 45-60 min. with a secondary anti-rabbit antibodies conjugated to peroxidase (dilution 1:10000) in TBST. Washing: as above. After washing blot was incubated 1-2 min. in ECL solution and exposed to Kodak autoradiography film. Exposure time was 15-60 seconds.

    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 ºC, 5 min.) and centrifuged (13000 rpm, 1 min.).

    Courtesy Dr. J. Piechota, University of Wroclaw, Poland

     

  • Additional Information
  • Additional information: Blue-native (2D BN/SDS-PAGE) methodology has been described in Piechota et al, 2010
  • Background
  • Background:

    One of the several classes of mitochondrial proteases is membrane bound, ATP dependent FtsH protease. Their function is very important for the control of protein quality and quantity by degradation of unassembled subunits. FtsH10 is localized in mitochondria. Alternative names: cell division protease ftsH homolog 10, mitochondrial, AtFtsH10

  • Product Citations
  • Selected references: Kolodziejczak et al. (2018). m-AAA Complexes Are Not Crucial for the Survival of Arabidopsis Under Optimal Growth Conditions Despite Their Importance for Mitochondrial Translation. Plant Cell Physiol. 2018 May 1;59(5):1006-1016. doi: 10.1093/pcp/pcy041.
    Piechota et al. (2015). Unraveling the functions of type II-prohibitins in Arabidopsis mitochondria. Plant Mol Biol. 2015 Apr 21.
    Kwasniak et al. (2013). Silencing of the Nuclear RPS10 Gene Encoding Mitochondrial Ribosomal Protein Alters Translation in Arabidiopsis Mitochondria. Plant Cell, May 30.
    Quesada et al. (2011). Arabidopsis RUGOSA2 encodes an mTERF family member required for mitochondrion, chloroplast and leaf development. Plant J.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts
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