Anti-MIF Antibody

Product Name

Anti-MIF Antibody

View all MIF Antibodies

SKU/Catalog Number

PA1052

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-MIF Antibody catalog # PA1052. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-MIF Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1052)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the C-terminus of human MIF, different from the related rat and mouse sequences by one amino acid.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PA1052 is reactive to MIF in Human, Mouse, Rat

Applications

PA1052 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

12 kDa

Calculated molecular weight

12.476kDa

Background of MIF

Macrophage migration inhibitory factor, MIF, is a cytokine released by T-lymphocytes, macrophages, and the pituitary gland that serves to integrate peripheral and central inflammatory responses. MIF gene has 3 exons separated by introns of only 189 and 95 bp, and covers less than 1 kb. Localization of the human gene for macrophage migration inhibitory factor (MIF) to chromosome 22q11.2 MIF plays a critical role in inflammatory diseases and atherogenesis.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse, Rat, By Heat
Flow Cytometry(Fixed), 1-3 μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of MIF using anti-MIF antibody (PA1052).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Jurkat whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: rat kidney tissue lysates,
Lane 4: rat NRK whole cell lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: mouse kidney tissue lysates,
Lane 7: mouse HBZY-1 whole cell lysates,
Lane 8: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MIF antigen affinity purified polyclonal antibody (Catalog # PA1052) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MIF at approximately 12 kDa. The expected band size for MIF is at 12 kDa.

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Figure 2. IHC analysis of MIF using anti-MIF antibody (PA1052).
MIF was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-MIF Antibody (PA1052) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. IHC analysis of MIF using anti-MIF antibody (PA1052).
MIF was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-MIF Antibody (PA1052) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of K562 cells using anti-MIF antibody (PA1052).
Overlay histogram showing K562 cells stained with PA1052 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MIF Antibody (PA1052, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For MIF (Source: Uniprot.org, NCBI)

Alternative Names

EC 5.3.2.1; EC 5.3.3.12; GIFmacrophage migration inhibitory factor; GLIF; Glycosylation-inhibiting factor; L-dopachrome isomerase; L-dopachrome tautomerase; macrophage migration inhibitory factor (glycosylation-inhibiting factor); MIF; MMIF; Phenylpyruvate tautomerase MIF GIF, GLIF, MMIF macrophage migration inhibitory factor macrophage migration inhibitory factor|L-dopachrome isomerase|L-dopachrome tautomerase|epididymis secretory sperm binding protein|macrophage migration inhibitory factor (glycosylation-inhibiting factor)|phenylpyruvate tautomerase

For more info on MIF, check out the MIF Infographic

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In this infographic, you will see the following information for MIF: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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