Anti-Cyclophilin F 抗体 [E11AE12BD4]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [E11AE12BD4] (AB110324)

Immunocytochemistry analysis using ab110324 at 1μg/ml staining Cyclophilin 40 in Cultured Human embryonic lung-derived fibroblasts (strain MRC5), (fixed, treated for heat-induced antigen retrieval, permeabilized) followed by an AlexaFluor® 488-conjugated-goat-anti-mouse IgG1 isotype specific secondary antibody (2 µg/ml).

• Flow Cyt

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Flow Cytometry - Anti-Cyclophilin F antibody [E11AE12BD4] (AB110324)

Flow cytometric analysis using ab110324 at 1µg/ml staining Cyclophilin F in HL60 cells (blue). Isotype control antibody (red).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [E11AE12BD4] (AB110324)

Immunohistological analysis using ab110324 at 1μg/ml staining Cyclophilin F in Human cerebellum tissue (Formalin-fixed, Paraffin-embedded).
Note : immunoactivity is most intense in neuronal cell bodies, most notably in the large Purkinje cells.

• WB

Lab

Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (AB110324)

Lanes 1- 2 : Merged signal (red and green). Green - ab110324 observed at 23 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

ab110324 was shown to react with Cyclophilin F in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266077 (knockout cell lysate ab257039) was used. Wild-type HEK-293T and PPIF knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab110324 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) at 1/10000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PPIF (Cyclophilin F) knockout HEK-293T cell lysate (<a href='/products/cell-lysates/human-ppif-cyclophilin-f-knockout-hek-293t-cell-lysate-ab257039'>ab257039</a>) at 20 µg

Predicted band size: 22 kDa

Observed band size: 37 kDa,23 kDa

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• WB

Lab

Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (AB110324)

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : Cyclophilin F knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : Hek293 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab110324 observed at 24 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab110324 detected the expected band for Cyclophilin F in wild type HAP1 cells and the band was not seen in Cyclophilin F knockout HAP1 cells. Wild-type and Cyclophilin F knockout samples were subjected to SDS-PAGE. ab110324 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

Predicted band size: 22 kDa

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• WB

AbReview49378****

Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (AB110324)

Blocked with 5% milk for 1 hour at 25°C.

Incubated with the primary antibody diluted in PBS-T + 5% milk for 16 hours at 4°C.

All lanes:

Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) at 1/1000 dilution

Lane 1:

WT mouse liver mitochondria lysate at 25 µg

Lane 2:

CypD KO mouse liver mitochondria lysate at 25 µg

Lane 3:

WT mouse liver mitochondria lysate at 35 µg

Lane 4:

CypD KO mouse liver mitochondria lysate at 35 µg

Lane 5:

WT mouse liver mitochondria lysate at 50 µg

Lane 6:

CypD KO mouse liver mitochondria lysate at 50 µg

Secondary

All lanes:

HRP-conjugated goat anti-mouse IgG polyclonal at 1/4000 dilution

Predicted band size: 22 kDa

Observed band size: 13 kDa,20 kDa

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Exposure time: 5s

This image is courtesy of an anonymous Abreview

• WB

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Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (AB110324)

All lanes:

Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) at 1 µg/mL

Lane 1:

Isolated mitochondria from Human heart at 5 µg

Lane 2:

Isolated mitochondria from Bovine heart at 1 µg

Lane 3:

Isolated mitochondria from Rat heart at 10 µg

Lane 4:

Isolated mitochondria from Mouse heart at 10 µg

Lane 5:

Isolated mitochondria from HepG2 cells at 20 µg

Predicted band size: 22 kDa

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