Anti-MIF antibody [EPR12463] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MIF antibody [EPR12463] - BSA and Azide free (AB183302)

Intracellular flow cytometric analysis of permeabilized Molt-4 cells using unpurified ab175189 at a 1/10 dilution (red) or a rabbit IgG (negative) (green).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-MIF antibody [EPR12463] - BSA and Azide free (AB183302)

Intracellular Flow Cytometry analysis of THP-1 cells labelling MIF with purified ab175189 at 1/140 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary anitbody. A rabbit monoclonal IgG was used as the isotype control (green).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-MIF antibody [EPR12463] - BSA and Azide free (AB183302)

Intracellular Flow Cytometry analysis of THP-1 cells labelling MIF with unpurified ab175189 at 1/50 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary anitbody. A rabbit monoclonal IgG was used as the isotype control (green).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

• IP

Lab

Immunoprecipitation - Anti-MIF antibody [EPR12463] - BSA and Azide free (AB183302)

ab175189 (unpurified) at 1/30 immunoprecipitating MIF in human fetal brain. For western blotting, a Peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

All lanes:

Immunoprecipitation - Anti-MIF antibody [EPR12463] (<a href='/en-us/products/primary-antibodies/mif-antibody-epr12463-ab175189'>ab175189</a>)

Predicted band size: 12 kDa

Observed band size: 12 kDa

false

• IP

Lab

Immunoprecipitation - Anti-MIF antibody [EPR12463] - BSA and Azide free (AB183302)

ab175189 (purified) at 1/50 immunoprecipitating MIF in human fetal brain. For western blotting, a Peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

All lanes:

Immunoprecipitation - Anti-MIF antibody [EPR12463] (<a href='/en-us/products/primary-antibodies/mif-antibody-epr12463-ab175189'>ab175189</a>)

Predicted band size: 12 kDa

Observed band size: 12 kDa

false

• WB

Supplier Data

Western blot - Anti-MIF antibody [EPR12463] - BSA and Azide free (AB183302)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : MIF knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : HepG2 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab175189 observed at 12 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab175189 was shown to specifically react with MIF in wild-type HAP1 cells along with additional cross-reactive bands. No bands were observed when knockout cells were examined. Wild-type and MIF knockout samples were subjected to SDS-PAGE. ab175189 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20,000 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 1/2,0000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

All lanes:

Western blot - Anti-MIF antibody [EPR12463] - BSA and Azide free (ab183302)

Predicted band size: 12 kDa

false