radio
SKU RP1020
Size 100μg/vial
Reactivity Human
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications ELISA, WB

Overview

Product Name Anti-MIF Antibody
SKU/Catalog Number RP1020
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.
Size 100μg/vial
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)
Host Rabbit
Immunogen E. coli-derived human MIF recombinant protein(Position: M1-A115).
Reactivity Human

Assay Details

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

/antibody/rp1020 1 WB anti mif antibody.jpg

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.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id P14174
Gene Name MIF
Protein Name Macrophage migration inhibitory factor
Alternative Names Macrophage migration inhibitory factor;MIF;5.3.2.1;Glycosylation-inhibiting factor;GIF;L-dopachrome isomerase;L-dopachrome tautomerase;5.3.3.12;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.

Ontology

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. .
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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Promotion:

Buy primary get secondary antibody for free.
Option Price
30ug sample size $99
100ug $240
100ug+Free HRP Secondary BA1054 $240
100ug+Free Biotin Secondary BA1003 $240
Ships in 5-7 business days.

Troubleshooting

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Customer Q&As

Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
A: You can find the immunogen sequence under "Immunogen" and clonality in the product name.
Q: What is the expected band size? Why is it different than the observed band size?
A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:
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.,
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
A: Check our protocols under the tech support tab.