Anti-MIF antibody [EPR18149-132] - BSA and Azide free (Capture)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MIF antibody [EPR18149-132] - BSA and Azide free (Capture) (AB242509)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C6 ( rat glial tumor glial cell) cells labelling MIF with ab242509 at 1/100 (10.45 µg/ml) dilution followed by a ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 (2 µg) dilution (Green).

Confocal image showing cytoplasmic and nuclear staining in C6 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5µg/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 (2 µg) dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MIF antibody [EPR18149-132] - BSA and Azide free (Capture) (AB242509)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling MIF with ab242509 at 1/100 (20.9 µg/ml) dilution followed by a ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 (2 µg) dilution (Green).

Confocal image showing cytoplasmic and nuclear staining in RAW 264.7 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5µg/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 (2 µg) dilution.

• WB

Lab

Western blot - Anti-MIF antibody [EPR18149-132] - BSA and Azide free (Capture) (AB242509)

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

ab181602 was used as a GAPDH loading control at a 1/200000 dilution.

In Western blot, ab242509 was shown to bind specifically to MIF. Target of interest was observed at 13 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in MIF knockout cell line (lane 2).

Exposure time : Lane 1-4 : 180 seconds
Lane 5-8 : 15 seconds

All lanes:

Western blot - Anti-MIF antibody [EPR18149-132] - BSA and Azide free (Capture) (ab242509) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

MIF knockout HAP1 whole cell lysate at 20 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 5:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 6:

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

Lane 7:

J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage ) whole cell lysate at 20 µg

Lane 8:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 13 kDa

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• sELISA

Supplier Data

Sandwich ELISA - Anti-MIF antibody [EPR18149-132] - BSA and Azide free (Capture) (AB242509)

Representative standard curve from corresponding SimpleStep ELISA® Kit (ab209885).