PPID KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
40 kDa peptidyl prolyl cis trans isomerase D, 40 kDa peptidyl-prolyl cis-trans isomerase, CYP-40, CyP-D, Cyclophilin D, Cyclophilin-40, Cyclophilin-related protein, MGC33096, PPID_HUMAN, PPIase, PPIase D, Peptidyl Prolyl Isomerase D, Peptidyl-prolyl cis-trans isomerase D, Rotamase, Rotamase D
PPID KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
We will provide viable cells that proliferate on revival.
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Lanes 1-3: Merged signal (red and green). Green - Anti-Cyclophilin 40 antibody [EPR14845(B)] ab181983 observed at 47 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-Cyclophilin 40 antibody [EPR14845(B)] ab181983 Anti-Cyclophilin 40 antibody [EPR14845(B)] was shown to specifically react with Cyclophilin 40 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265788 (knockout cell lysate Human PPID (Cyclophilin 40) knockout HeLa cell lysate ab257600) was used. Wild-type and Cyclophilin 40 knockout samples were subjected to SDS-PAGE. Anti-Cyclophilin 40 antibody [EPR14845(B)] ab181983 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Cyclophilin 40 antibody [EPR14845(B)] (Anti-Cyclophilin 40 antibody [EPR14845(B)] ab181983) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: PPID knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human PPID (Cyclophilin 40) knockout HeLa cell line (ab265788)
Lane 3: K-562 cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 41 kDa
Observed band size: 47 kDa
Homozygous: 1 bp insertion in exon 1.
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