Antibodies, Proteins, Kits and Reagents for Life Science

338 390 ₸ Product size

100 µl 338 390 ₸

Order now and get it on Tuesday March 02, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR12463] to MIF
Suitable for: WB, IP, Flow Cyt
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-MIF antibody [EPR12463]
See all MIF primary antibodies
Description

Rabbit monoclonal [EPR12463] to MIF
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide within Human MIF aa 50 to the C-terminus (Cysteine residue). The exact sequence is proprietary.
Database link: P14174
Positive control
- WB: Y79, THP-1, and Molt-4 cell lysates. Mouse and rat brain tissue lysates. Flow Cyt: THP-1, and Molt-4 cells. IP: Human fetal brain tissue lysate.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR12463
Isotype

IgG
Research areas

Western blot - Anti-MIF antibody [EPR12463] (ab175189)

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.
Flow Cytometry - Anti-MIF antibody [EPR12463] (ab175189)

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).
Western blot - Anti-MIF antibody [EPR12463] (ab175189)

All lanes : Anti-MIF antibody [EPR12463] (ab175189) at 1/1000 dilution (unpurified)

Lane 1 : Y79 cell lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Rat brain tissue lysate

Lysates/proteins at 20 µg/ml per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 12 kDa
Observed band size: 12 kDa

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-MIF antibody [EPR12463] (ab175189)

All lanes : Anti-MIF antibody [EPR12463] (ab175189) at 1/2000 dilution (purified)

Lane 1 : Y79 cell lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Rat brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 12 kDa
Observed band size: 12 kDa

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-MIF antibody [EPR12463] (ab175189)

All lanes : Anti-MIF antibody [EPR12463] (ab175189) at 1/1000 dilution (unpurified)

Lane 1 : Y79 lysates
Lane 2 : THP-1 lysates
Lane 3 : Molt-4 lysates
Lane 4 : Human fetal brain lysates

Lysates/proteins at 10 µg per lane.

Predicted band size: 12 kDa
Flow Cytometry - Anti-MIF antibody [EPR12463] (ab175189)

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).
Flow Cytometry - Anti-MIF antibody [EPR12463] (ab175189)

Flow cytometric analysis of permeabilized Molt-4 cells using unpurified ab175189 at a 1/10 dilution (red) or a rabbit IgG (negative) (green).
Immunoprecipitation - Anti-MIF antibody [EPR12463] (ab175189)

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.
Immunoprecipitation - Anti-MIF antibody [EPR12463] (ab175189)

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.
Anti-MIF antibody [EPR12463] (ab175189)

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