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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIF antibody [EPR18149-128] - BSA and Azide free (AB226166)

This data was developed using ab187064, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of Paraffin-embedded sections on Human kidney tissue with ab187064 at 1/2500 dilution, followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human kidney. Counter stained with Hematoxylin.

The primary antibody was incubated for 30 mins at room temperature. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIF antibody [EPR18149-128] - BSA and Azide free (AB226166)

This data was developed using ab187064, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of Paraffin-embedded sections on Rat kidney tissue with ab187064 at 1/2500 dilution, followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat kidney. Counter stained with Hematoxylin.

The primary antibody was incubated for 30 mins at room temperature. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIF antibody [EPR18149-128] - BSA and Azide free (AB226166)

This data was developed using ab187064, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of Paraffin-embedded sections on Mouse kidney tissue with ab187064 at 1/2500 dilution, followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse kidney. Counter stained with Hematoxylin.

The primary antibody was incubated for 30 mins at room temperature. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• 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).

• 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).

• 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).

• 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='/ko/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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIF antibody [EPR18149-128] - BSA and Azide free (AB226166)

This data was developed using ab187064, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of Paraffin-embedded sections on A : Wild-type HAP1 (chronic myelogenous leukemia cell) cell pellet and B : MIF knockout HAP1 cell pellet, with ab187064 at 1/2500 dilution, followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Wild-type HAP1 (chronic myelogenous leukemia cell) cell pellet, no staining on (B) MIF knockout HAP1 cell pellet. Counter stained with Hematoxylin.

The primary antibody was incubated for 30 mins at room temperature. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.