Anti-MIF antibody [EPR18149-128] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MIF antibody [EPR18149-128] - BSA and Azide free (AB226166)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling MIF with ab187064 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Neuro-2a cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody 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 (ab187064).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MIF antibody [EPR18149-128] - BSA and Azide free (AB226166)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Neuro-2a (mouse neuroblastoma cell line) cell line labeling MIF with ab187064 at 1/500 (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). 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 (ab187064).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MIF antibody [EPR18149-128] - BSA and Azide free (AB226166)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling MIF with ab187064 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on RAW 264.7 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody 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 (ab187064).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MIF antibody [EPR18149-128] - BSA and Azide free (AB226166)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell line labeling MIF with ab187064 at 1/500 (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). 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 (ab187064).

• WB

Lab

Western blot - Anti-MIF antibody [EPR18149-128] - BSA and Azide free (AB226166)

ab187064 was shown to specifically react with MIF in wild-type HAP1 cells as signal was lost in MIF knockout cells. Wild-type and MIF knockout samples were subjected to SDS-PAGE. ab187064 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (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 instrument using the ECL technique.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187064).

All lanes:

Western blot - Anti-MIF antibody [EPR18149-128] (<a href='/en-us/products/primary-antibodies/mif-antibody-epr18149-128-ab187064'>ab187064</a>) at 1/1000 dilution

Lane 1:

Wild type HAP1 whole cell lysate at 20 µg/mL

Lane 2:

MIF knockout HAP1 whole cell lysate at 20 µg/mL

Lane 3:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg/mL

Lane 4:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg/mL

Predicted band size: 12 kDa

Observed band size: 13 kDa

false

Exposure time: 15s