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. Synonyme: cell division protease ftsH homolog 4, mitochondrial

KLH-conjugated peptide derived from sequence of Arabidopsis thaliana FtsH4 O80983

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)

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

to be added when available

application information

1: 500 with standard ECL (WB)

77 | 72 kDa

Arabidopsis thaliana

Arabidopsis thaliana

Solanum lycopersicum

to be added when available

Gibala et al. (2009). The lack of mitochondrial AtFtsH4 protease alters Arabidopsis leaf morphology at the late stage of rosette development under short day photoperiod. Plant J.

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