Anti-Mitofilin antibody [EPR8749] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mitofilin antibody [EPR8749] - BSA and Azide free (AB245764)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human hepatocellular cancer tissue sections labeling Mitofilin with purified ab137057 at 1/250 dilution (0.508 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137057).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mitofilin antibody [EPR8749] - BSA and Azide free (AB245764)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human thyroid cancer tissue sections labeling Mitofilin with purified ab137057 at 1/250 dilution (0.508 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137057).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Mitofilin antibody [EPR8749] - BSA and Azide free (AB245764)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Mitofilin with purified ab137057 at 1/100 dilution (1.3 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137057).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mitofilin antibody [EPR8749] - BSA and Azide free (AB245764)

Immunohistochemical analysis of paraffin embedded Human colon tissue labelling Mitofilin with ab137057 at 1/100 dilution.This data was developed using the same antibody clone in a different buffer formulation containing PBS#44; BSA#44; glycerol and sodium azide (ab137057)

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Mitofilin antibody [EPR8749] - BSA and Azide free (AB245764)

Immunofluorescent staining of HeLa cells labelling Mitofilin with ab137057 at 1/250 dilution.This data was developed using the same antibody clone in a different buffer formulation containing PBS#44; BSA#44; glycerol and sodium azide (ab137057)

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Mitofilin antibody [EPR8749] - BSA and Azide free (AB245764)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Mitofilin with purified ab137057 at 1/20 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137057).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mitofilin antibody [EPR8749] - BSA and Azide free (AB245764)

Immunohistochemical analysis of paraffin embedded Human testis tissue labelling Mitofilin with ab137057 at 1/100 dilution.This data was developed using the same antibody clone in a different buffer formulation containing PBS#44; BSA#44; glycerol and sodium azide (ab137057)

• WB

Lab

Western blot - Anti-Mitofilin antibody [EPR8749] - BSA and Azide free (AB245764)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137057).

Lanes 1 - 4 : Merged signal (red and green). Green - ab137057 observed at 84 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab137057 was shown to specifically react with IMMT in wild-type HAP1 cells as signal was lost in IMMT knockout cells. Wild-type and IMMT knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab137057 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution 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-Mitofilin antibody [EPR8749] - BSA and Azide free (ab245764) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

IMMT knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HepG2 whole cell lysate at 20 µg

Lane 4:

Human Heart whole cell lysate at 20 µg

Predicted band size: 84 kDa

Observed band size: 84 kDa

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