| product information
| | background | | 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. | | immunogen | | | | antibody format | | | | quantity | | 250µl, Lyophilized | for reconstitution add225 µl of sterile water. | | | storage | | 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. | | tested applications | | western blot (WB) | | related products | | AS07 205 | anti-FtsH4 | plant fts metallo-protease H4 AS07 251 | anti-FtsH10 | plant fts metallo-protease H10 | | additional information | | TheFtsH2 protein standard can be used in combination with anti-FtsH2 antibodies to quantitateFtsH2 from a range of cyanobacteria. Global antibodies are raised against highly conserved amino acid sequences in theIsiA protein. Quantitative western blot: detailed method description, video tutorial | |
application information
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| recommended dilution | | Standard curve: 3 loads are recommended (0.5, 2 and 4μl).
For most applications a sample load of 0.2μg of chlorophyll will give aFtsH2 signal in this range.
Positive control: load per well: a 2μl load is optimal for most chemiluminescent detection systems.
This standard is ready-to-load and does not require any additions or heating. |
| expected | apparent MW | | A. thaliana: FtsH1:76.7, FtsH2: 74, FtsH5: 75.2, FtsH6: 74.5, FtsH7: 87.8, FtsH8: 73, FtsH9:87.8 kDa (chloroplastic); FtsH3: 89.3, FtsH4: 77.2, FtsH10: 89.5 kDa, FtsH11:88.7 kDa (mitochondrial) |
| confirmed reactivity | | |
| predicted reactivity | | |
| not reactive in | | |
| additional information | | Concentration: after adding 225 µl of sterile milliQ water final concentration of the standard is 0.1 pmoles/µl
For detection on a diatom samples, load of 5-10 µg/well is required as well as usage of enchanced chemiluminescence (for example ECL Advance). The antibody also detects the chloroplast-encoded FtsH from Thalassiosira pseudonana (diatom) along with 3 related isoforms encode in the nuclear genome of Thalassiosira pseudonana. |
| selected references | | to be added when available, antibody released in October 2011. |
application example
8 µg of total protein from Arabidopsis thaliana wild type (1) and FtsH 1 and 3 deletion mutant (2) extracted with 0.2M Tris-HCl pH6.8, 2%SDS, 10% mercaptoehanol, 5M Urea were separated on 12 % SDS-PAGE and blotted 2h to NC. Blots were blocked with non fat milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2 000 over night at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 5 min and 2 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody anti-rabbit IgG horse radish peroxidase conjugated, from Sigma) diluted to 1:10 000 in TBS-T+5% non fat milk for 2h at RT with agitation.
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