Plant/Algal cell antibodies / Mitochondria | Respiration

product information

One of the several classes of mitochondrial proteases is membrane bound, ATPdependent FtsH protease. Their function is very important for the control of protein quality and quantity by degradation of unassembled subunits. Other names: AtFtsH3, cell division protease ftsH homolog 3, mitochondrial, AtFtsH10, cell division protease ftsH homolog 10, mitochondrial

KLH-conjugated peptide dereived from sequences of Arabidopsis thaliana FtsH3 and FtsH10 with localization to mitochondria Q84WU8 and Q8VZI8

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.

western blot (WB), Blue-native (2D BN/SDS-PAGE)

AS07 205 | anti-FtsH4 | plant fts metallo-protease H4

AS07 251 | anti-FtsH10 | plant fts metallo-protease H10

Blue-native (2D BN/SDS-PAGE) methodology is described in Piechota et al. 2010

application information

1 : 500- 1: 1000 with standard ECL (WB)

80 kDa

Arabidopsis thaliana

Arabidopsis thaliana

no confirmed exceptions from predicted reactivity known in the moment

to be added when available

Piechota et al. (2010). Identification and characetization of high-molecular-weight complexes fromed by m-AAA proteases and prohibitins in mitochondria of Arabidopsis thaliana. In press.

application example

total protein from Arabidopsis thaliana mitochondria (20 µg) were 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 (Sigma, dilution 1:10000) in TBST. Washing: as above. After washing blot was incubated 1-2 min. in ECL solution and exposed to Kodakautoradiography 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 (13000rpm, 1 min.).

Courtesy Dr. J. Piechota