FtsH2 | FtsH2 positive control/quantitation standard

FtsH belong to a family of ATP dependent peptidases. Localized in a chloroplast are following isoforms: FTSH1 (synonymes AAA, FTSH, FTSH Protease 1), Ftsh2 (VAR2, VARIEGATED 2), FtsH5 (VAR1, VARIEGATED 1), FtsH6 (FTSH PROTEASE 6), FtsH7, FtsH8. FtsH9.Localized in mitochondria are following isoforms: FtsH3 (FTSH Protease 3), FtsH4, FtsH10, FtsH11.

Standard curve: 3 loads are recommended (0.5, 2 and 4 μl) .For most applications a sample load of 0.2μg of chlorophyll will give a FtsH2 signal in this range.Positive control: load per well: a 2 μl load is optimal for most chemiluminescent detection systems.Standard curve: 3 loads are recommended (0.5, 2 and 4 µl) .For most applications a sample load of 0.2 µg of chlorophyll will give a FtsH2 signal in this range.Positive control: a 2 µl load per well is optimal for most chemiluminescent detection systems.Non-disulphie dependent dimers and complexes can be also detected using standard western blot methods withmore sensitive detection reagents as ECL Advance or West Pico when loading per well more standard than recommended. They have not been included in the standard calibration.This standard is stabilized and ready and does not require heating before loading on the gel.Please note that this product contains 10% glycerol and might appear as liquid but is provided lyophilized. Allow the product several minutes to solubilize after adding water. Mix thoroughly but gently Take extra care to mix thoroughly before each use, as the proteins tend to settle with the more dense layer after freezing.

Concentration:after adding 225 µl of sterile milliQ water final concentration of the standard is 0.1 pmoles/µLProtein standard buffer composition: Glycerol 10%, Tris Base 141 mM, Tris HCl 106 mM, LDS 2%, EDTA 0.51mM, SERVA® Blue G250 0.22 mM, Phenol Red 0.175 mM, pH 8.5, 0.1 mg/mL PefaBloc protease inhibitor (Roche), 50mM DTT.This standard is ready-to-load and does not require any additions or heating. It needs to be fully thawed and thoroughly mixed prior to using. Avoid vigorous vortexing, as buffers contain detergent. Following mixing, briefly pulse in a microcentrifuge to collect material from cap.This standard is stabilized and ready and does not require heating before loading on the gel.Please note that this product contains 10% glycerol and might appear as liquid but is provided lyophilized.Allow the product several minutes to solubilize after adding water. Mix thoroughly but gently Take extra careto mix thoroughly before each use, as the proteins tend to settle with the more dense layer after freezing,

The FtsH2 protein standard can be used in combination with anti-FtsH2 antibodies AS11 1789 to quantitate FtsH2 from a range of cyanobacteria. Global antibodies are raised against highly conserved amino acid sequences in the FtsH protein.Quantitative western blot: detailed method description, video tutorial

Store lyophilized/reconstituted at -20°C; 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.