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SKU:RP1020
Pack Size:100μg/vial
Sample Size:30ug for $99, contact us for details
Clonality:Polyclonal
Application:ELISA, WB
Price: $240.00
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Overview

Product Name Anti-MIF Antibody
SKU/Catalog Number RP1020
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)
Replacement Item This antibody may replace the following items: sc-16966|sc-16965|sc-48241|sc-20121|sc-271631|sc-47159|sc-53915|sc-130329 from Santa Cruz Biotechnology.
Host Rabbit
Isotype N/A
Validated Species Human
Application ELISA, WB

*Our Boster Guarantee covers the use of this product in the above tested applications.

**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.

Recommended Detection Systems Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2017!
Immunogen E. coli-derived human MIF recombinant protein(Position: M1-A115).
Cross Reactivity No cross reactivity with other proteins
Pack Size 100μg/vial

Properties

Clonality Polyclonal
Form Lyophilized
Contents 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.
Concentration Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Storage 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.
Purification Immunogen affinity purified.
Isotype N/A

Protein Target Info (Source: Uniprot.org)

You can check the tissue specificity below for information on selecting positive and negative control.

Gene Name MIF
Protein Name Macrophage migration inhibitory factor
Molecular Weight 12476 MW
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. .
Sequence Similarities Belongs to the MIF family.
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.
Uniprot ID P14174
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;
Research Areas |immunology|innate immunity|macrophage / inflamm.|
*if product is indicated to react with multiple species, protein info is based on the human gene.

Background for Macrophage migration inhibitory factor

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.

Anti-MIF Antibody Images

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Anti-MIF Antibody
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.
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FAQs

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.