Overview

  • Product name
    Human MIF ELISA Kit
  • Detection method
    Colorimetric
  • Sample type
    Cell culture supernatant, Serum, Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    < 6 pg/ml
  • Range
    8.23 pg/ml - 6000 pg/ml
  • Recovery

    94 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 94.68 83% - 103%
    Serum 93.48 82% - 102%
    Plasma 95.41 84% - 103%

  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Abcam’s MIF Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human MIF in serum, plasma (collect plasma using heparin as an anticoagulant. EDTA and Citrate are not recommended), and cell culture supernatants.

    This assay employs an antibody specific for Human MIF coated on a 96-well plate. Standards and samples are pipetted into the wells and MIF present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human MIF antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of MIF bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm

  • Notes

    Optimization may be required with urine samples.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    300X HRP-Streptavidin Concentrate 1 x 200µl
    5X Assay Diluent B 1 x 15ml
    Assay Diluent A 1 x 30ml
    Biotinylated anti-Human MIF 2 vials
    MIF Microplate (12 x 8 wells) 1 unit
    Recombinant Human MIF Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas
  • Relevance
    MIF is a proinflammatory cytokine involved in many inflammatory reactions and disorders. MIF (macrophage migration inhibitory factor) was one of the first cytokines to be discovered and was initially described as a T cell-derived factor that inhibits the random migration of macrophages (Weiser 1989). Recently, MIF was rediscovered as a pituitary hormone that act as the counterregulatory hormone for glucocorticoid action within the immune system (Bernhagen 1993, Mitchell 1995). MIF was released from macrophages and T cells in response to physiological concentrations of glucocorticoids. The secreted MIF counter-regulates the immunosuppressive effects of steroids on immune cell activation and cytokine production (Bucala 1998). MIF plays a critical role in the host control of inflammation and immunity. MIF is involved in both autoimmune disorders and tumorigenesis.
  • Cellular localization
    Secreted. Cytoplasm. Note: 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.
  • Alternative names
    • GIF
    • GLIF
    • Glycosylation inhibiting factor
    • L dopachrome isomerase
    • L dopachrome tautomerase
    • Macrophage migration inhibitory factor
    • Macrophage migration inhibitory factor (glycosylation inhibiting factor)
    • MMIF
    • Phenylpyruvate tautomerase
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab100594 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • MIF in THP1 supernatants from control cells or cells stimulated (P/L) for 24 hours with 50 ng x mL-1 PMA (ab120297) and 1 ug x mL-1 LPS for the last 6 hours. Samples tested in the range of 1/3-1/30 (duplicates, +/- SD).

  • MIF measured in cell culture supernatants from control or treated (PHA: 2 days in 2% PHA-M; LifeTechnologies) PBMCs. Samples tested in the range of 1/3-1/30 (duplicates, +/- SD).

  • MIF in human biological fluids (duplicates +/- SD). Data shown from undiluted serum, plasma and urine, and milk diuted 1/100-1/500. Mouse serum gave 0.12 ng/mL (duplicates) and no signal was detected in mouse plasma.

  • Standard curve in assay buffer B with background signal subtracted (duplicates; +/- SD).

  • Representative Standard Curve using ab100594.

  • Representative Standard Curve using ab100594.

Protocols

References

ab100594 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question
Answer


In general, 1% SDS is higher than we advise and urea at a high enough concentration may also denature proteins. Depending the samples, you may still have a chance of getting some detection but 1% SDS and urea is not ideal. The only way to know for certain though would be to test.

If you decide to test this kit with lysate samples, we recommend diluting the samples at least 5-fold with 1X Assay Diluent B to minimize any effects of the detergents in the lysis buffer. The samples may need to be diluted further but this would need to be determined empirically.

For the original lysate, in general you would want to aim for around 1 mg/ml protein, preferably more, to achieve 50-500 ug/ml after dilution. Again though, this range should just be used as a starting point and the optimal loading concentration would need to be determined.

In brief, a compatible lysis buffer must meet the following specifications:
A) has relatively low salt content (700 mM or less)
B) does not contain sodium azide
C) does not contain >0.1% SDS
D) does not contain >10 mM reducing agents (beta-mercaptoethanol or dithiothreitol)

This would include any buffers used for immunoprecipitations, including RIPA buffer.

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Question
Answer

Thank you for contacting us. I have checked the homology and it's 90% between human and mouse MIF protein. Unfortunately, there isn’t any cross reactivity data available with the ab100594 kit and mouse samples. If it helps though, the capture and detection antibody have both been separately tested in a western blot application for reactivity with mouse and the results showed ~25% cross reactivity and will detect mouse MIF, respectively. However, whether these antibodies functioning as a pair like in ab100594 will detect mouse is not known so the only way to find out would be to test. A 90% homology is fairly high so there might be a chance. However, unfortunately, we might not be able to guarantee the kit for mouse samples as we have so far no data. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us. The capture antibody in our MIF Human ELISA Kit ab100594 is a monoclonal mouse IgG1 and the detection antibody is a polyclonal goat IgG. Unfortunately, we have no information about whether this kit would be predicted to cross-react with horse. I hope this helps, please let me know if you need any additional information.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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