Antibodies, Proteins, Kits and Reagents for Life Science

291 484 ₸ Product size

100 µl 291 484 ₸ 10 µl 120 614 ₸

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Anti-MIF antibody [EPR18149-128] (ab187064)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18149-128] to MIF
Suitable for: Flow Cyt, ICC/IF, WB
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-MIF antibody [EPR18149-128]
See all MIF primary antibodies
Description

Rabbit monoclonal [EPR18149-128] to MIF
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC/IF	Mouse
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal brain and fetal spleen lysates; Mouse brain and kidney lysate; Rat brain lysate; J774A.1, Y79, Jurkat, NIH/3T3, RAW 264.7, C6 and Neuro-2a whole cell lysates. ICC/IF: Neuro-2a and RAW 264.7 cells. Flow Cyt: Neuro-2a and RAW 264.7 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18149-128
Isotype

IgG
Research areas

Western blot - Anti-MIF antibody [EPR18149-128] (ab187064)

All lanes : Anti-MIF antibody [EPR18149-128] (ab187064) at 1/1000 dilution

Lane 1 : Wild type HAP1 whole cell lysate
Lane 2 : MIF knockout HAP1 whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
Lane 4 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate

Lysates/proteins at 20 µg/ml per lane.

Predicted band size: 13 kDa
Observed band size: 13 kDa

Exposure time: 15 seconds

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.
Western blot - Anti-MIF antibody [EPR18149-128] (ab187064)

All lanes : Anti-MIF antibody [EPR18149-128] (ab187064) at 1/1000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal spleen lysate
Lane 3 : Mouse brain lysate
Lane 4 : Rat brain lysate
Lane 5 : J774A.1 (mouse reticulum cell sarcoma monocyte macrophage cell line) whole cell lysate
Lane 6 : Mouse kidney lysate
Lane 7 : Y79 (human retinoblastoma cell line) whole cell lysate
Lane 8 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 9 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
Lanes 1-2 & 7-8 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Lanes 3-6 & 9 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 13 kDa
Observed band size: 13 kDa

Exposure time : Lanes 1,2,3 and 4: 10 seconds; Lanes 5 and 6: 3 seconds; Lanes 7,8 and 9: 1 second.

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-MIF antibody [EPR18149-128] (ab187064)

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.
Flow Cytometry - Anti-MIF antibody [EPR18149-128] (ab187064)

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.
Western blot - Anti-MIF antibody [EPR18149-128] (ab187064)

All lanes : Anti-MIF antibody [EPR18149-128] (ab187064) at 1/5000 dilution

Lane 1 : NIH/3T3 (mouseembryo fibroblast cell line) whole cell lysate
Lane 2 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : C6 (rat glial tumor cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 13 kDa
Observed band size: 13 kDa

Exposure time : Lane 1: 5 seconds; Lane 2: 1 second; Lane 3: 30 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-MIF antibody [EPR18149-128] (ab187064)

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.
Flow Cytometry - Anti-MIF antibody [EPR18149-128] (ab187064)

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.
Anti-MIF antibody [EPR18149-128] (ab187064)

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