|Product Name||Anti-MIF Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Macrophage migration inhibitory factor(MIF) detection. Tested with WB, ELISA in Human.|
|Cite This Product||Anti-MIF Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1020)|
|Contents/Buffer||Each vial contains 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3. Carrier free (No BSA) form available in stock. If you want this antibody carrier free please specify "Carrier Free" or "No BSA" in your order note.|
|Immunogen||E. coli-derived human MIF recombinant protein(Position: M1-A115).|
Assay Dilutions Overview
Western blot, 0.1-0.5μg/ml, Human, -
ELISA , 0.1-0.5μg/ml, Human
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.
Images And Assay Conditions
Figure. Western blot analysis of MIF using anti- MIF antibody (RP1020).
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 50ug of sample under reducing conditions.
Lane : Recombinant Human MIF Protein 0.5ng
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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 # RP1020) 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 1:10000 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 12KD. The expected band size for MIF is at 12KD.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Macrophage migration inhibitory factor|
|Alternative Names||Macrophage migration inhibitory factor;MIF;188.8.131.52;Glycosylation-inhibiting factor;GIF;L-dopachrome isomerase;L-dopachrome tautomerase;184.108.40.206;Phenylpyruvate tautomerase;MIF;GLIF, MMIF;|
|Subcellular Localization||Secreted. Cytoplasm. Does not have a cleavable signal sequence and is secreted via a specialized, non- classical pathway. Secreted by macrophages upon stimulation by bacterial lipopolysaccharide (LPS), or by M.tuberculosis antigens.|
|Molecular Weight||12476 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Pro-inflammatory cytokine. Involved in the innate immune response to bacterial pathogens. The expression of MIF at sites of inflammation suggests a role as mediator in regulating the function of macrophages in host defense. Counteracts the anti- inflammatory activity of glucocorticoids. Has phenylpyruvate tautomerase and dopachrome tautomerase activity (in vitro), but the physiological substrate is not known. It is not clear whether the tautomerase activity has any physiological relevance, and whether it is important for cytokine activity. .|
|Research Areas||Immunology, Innate Immunity, Macrophage / Inflamm.
*You can search these to find other products in these research areas.
|Background||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 1kb. 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.|
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1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,