Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free

10 product images

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  Western blot - Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free (ab231287)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Cyclophilin F antibody [EPR11311-121] ab231155).
  

  Lanes 1- 2: Merged signal (red and green). Green - Anti-Cyclophilin F antibody [EPR11311-121] ab231155 observed at 23 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

  Anti-Cyclophilin F antibody [EPR11311-121] ab231155 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 Human PPIF (Cyclophilin F) knockout HEK-293T cell line ab266077 (knockout cell lysate Human PPIF (Cyclophilin F) knockout HEK-293T 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. Anti-Cyclophilin F antibody [EPR11311-121] ab231155 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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 F antibody [EPR11311-121] (Anti-Cyclophilin F antibody [EPR11311-121] ab231155) at 1/1000 dilution

  Lane 1: Wild-type HEK-293T cell lysate at 20 µg

  Lane 2: PPIF knockout HEK-293T cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 22 kDa

  Observed band size: 23 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free (ab231287)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Cyclophilin F with Anti-Cyclophilin F antibody [EPR11311-121] ab231155 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 (green). Confocal image showing Cyclophilin F staining and mitochondrial localization in the HeLa cell line. The nuclear counter stain is DAPI (blue).

  Mitochondria are stained with Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 AlexaFluor^®594 Goat anti-Mouse secondary both at 1/1000 dilution (red).

  -ve control 1: Anti-Cyclophilin F antibody [EPR11311-121] ab231155 (Rabbit anti-Cyclophilin F) at 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

  -ve control 2: Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (AlexaFluor®488 Goat anti-Rabbit secondary) at 1/1000 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cyclophilin F antibody [EPR11311-121] ab231155).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free (ab231287)

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Cyclophilin F with Anti-Cyclophilin F antibody [EPR11311-121] ab231155 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in mouse colon (PMID: 23991223, PMID: 15344907, PMID: 26258444) is observed. Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cyclophilin F antibody [EPR11311-121] ab231155).

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  Flow Cytometry (Intracellular) - Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free (ab231287)

  Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Jurkat (human T cell leukemia T lymphocyte) cell line labeling Cyclophilin F with Anti-Cyclophilin F antibody [EPR11311-121] ab231155 at 1/600 (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), at 1/2000 dilution was used as the secondary antibody.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cyclophilin F antibody [EPR11311-121] ab231155).

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  Western blot - Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free (ab231287)

  Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
  

  Molecular mass is consistent with literature (PMID 1744118).

  Anti-Cyclophilin F antibody [EPR11311-121] ab231155 was shown to specifically react with Cyclophilin F in wild-type HAP1 cells as signal was lost in Cyclophilin F knockout cells. Wild-type and Cyclophilin F knockout samples were subjected to SDS-PAGE. Anti-Cyclophilin F antibody [EPR11311-121] ab231155 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Human anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/200000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrumentusing the ECL technique.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cyclophilin F antibody [EPR11311-121] ab231155).

  All lanes: Western blot - Anti-Cyclophilin F antibody [EPR11311-121] (Anti-Cyclophilin F antibody [EPR11311-121] ab231155) at 1/1000 dilution

  Lane 1: Wild type HAP1 whole cell lysate at 20 µg

  Lane 2: Cyclophilin F knockout HAP1 whole cell lysate at 20 µg

  Lane 3: HEK-293 (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

  Lane 4: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 22 kDa

  Observed band size: 18 kDa

  Exposure time: 59s

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free (ab231287)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Wildtype and Cyclophilin F knockout HAP1 (human chronic myelogenous leukemia near-haploid cell line) cells labeling Cyclophilin F with Anti-Cyclophilin F antibody [EPR11311-121] ab231155 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 (green). Confocal image showing Cyclophilin F staining and mitochondrial co-localization in wildtype HAP1 cells, with loss of the signal in the Cyclophin F-knockout HAP1 cells. The nuclear counter stain is DAPI (blue).

  Mitochondria are stained with with Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 AlexaFluor^®594 Goat anti-Mouse secondary both at 1/1000 dilution (red).

  -ve control 1: Anti-Cyclophilin F antibody [EPR11311-121] ab231155 (Rabbit anti-Cyclophilin F) at 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor^®594 Goat anti-Mouse secondary) at 1/1000 dilution.

  -ve control 2: Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (AlexaFluor^®488 Goat anti-Rabbit secondary) at 1/1000 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cyclophilin F antibody [EPR11311-121] ab231155).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free (ab231287)

  Immunohistochemical analysis of paraffin-embedded human colonic carcinoma tissue labeling Cyclophilin F with Anti-Cyclophilin F antibody [EPR11311-121] ab231155 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in tumor cells of human colon carcinoma (PMID: 23991223, PMID: 15344907, PMID: 26258444) is observed. Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cyclophilin F antibody [EPR11311-121] ab231155).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free (ab231287)

  Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Cyclophilin F with Anti-Cyclophilin F antibody [EPR11311-121] ab231155 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in human kidney (PMID: 23991223, PMID: 15344907, PMID: 26258444) is observed. Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cyclophilin F antibody [EPR11311-121] ab231155).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free (ab231287)

  Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling Cyclophilin F with Anti-Cyclophilin F antibody [EPR11311-121] ab231155 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in human pancreas (PMID: 23991223, PMID: 15344907, PMID: 26258444) is observed. Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cyclophilin F antibody [EPR11311-121] ab231155).

• 

  Flow Cytometry (Intracellular) - Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free (ab231287)

  Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling Cyclophilin F with Anti-Cyclophilin F antibody [EPR11311-121] ab231155 at 1/600 (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), at 1/2000 dilution was used as the secondary antibody.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cyclophilin F antibody [EPR11311-121] ab231155).