Mouse MIF ELISA Kit (ab209885)
- Datasheet
- References
- Protocols
Overview
-
Product name
Mouse MIF ELISA Kit
See all MIF kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Mouse serum 5 13.6% Inter-assay Sample n Mean SD CV% Mouse serum 3 9.8% -
Sample type
Cell culture supernatant, Serum, Cell culture extracts, Tissue Extracts, Hep Plasma, EDTA Plasma, Cit plasma, Tissue Homogenate -
Assay type
Sandwich (quantitative) -
Sensitivity
28 pg/ml -
Range
125 pg/ml - 8000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Serum 110 106% - 117% Cell culture extracts 111 108% - 113% Tissue Extracts 108 102% - 118% Hep Plasma 112 105% - 117% EDTA Plasma 109 105% - 112% Cit plasma 112 112% - 113% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Mouse -
Product overview
Abcam’s MIF in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of MIF protein in mouse serum, plasmas, cell culture supernatants, and cell and tissue extracts.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Sensitivity:
Samples diluted in Sample Diluent NS: 40 pg/mL
Samples diluted in 1X Cell Extraction Buffer PTR: 28 pg/mL
-
Notes
Macrophage migration inhibitory factor (MIF) is a 12.37 kDa pro-inflammatory cytokine and is involved in the innate immune response to bacterial pathogens. MIF assembles into a trimer, composed of three identical subunits. The expression of MIF at sites of inflammation suggests a role as mediator in regulating the function of macrophages in host defense. MIF counteracts the anti-inflammatory activity of glucocorticoids and exhibits phenylpyruvate tautomerase and dopachrome tautomerase activity. It is not clear whether the tautomerase activity has any physiological relevance, and whether it is important for cytokine activity.
-
Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Microplate (12 x 8 well strips)
Properties
-
Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Mouse MIF Capture Antibody 1 x 600µl 10X Mouse MIF Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BR 1 x 6ml Mouse MIF Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
-
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. -
Involvement in disease
Genetic variations in MIF are associated with susceptibility to rheumatoid arthritis systemic juvenile (RASJ) [MIM:604302]. An inflammatory articular disorder with systemic-onset beginning before the age of 16. It represents a subgroup of juvenile arthritis associated with severe extraarticular features and occasionally fatal complications. During active phases of the disorder, patients display a typical daily spiking fever, an evanescent macular rash, lymphadenopathy, hepatosplenomegaly, serositis, myalgia and arthritis. -
Sequence similarities
Belongs to the MIF family. -
Cellular 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. - Information by UniProt
-
Alternative names
- GIF
- GLIF
- Glycosylation inhibiting factor
see all -
Database links
- Entrez Gene: 17319 Mouse
- SwissProt: P34884 Mouse
- Unigene: 2326 Mouse
Associated products
-
ELISA kits
Applications
Our Abpromise guarantee covers the use of ab209885 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
-
Other - Mouse MIF ELISA Kit (ab209885)SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
-
Example of mouse MIF standard curve in Sample Diluent NSBackground-subtracted data values (mean +/- SD) are graphed.
-
Example of mouse MIF standard curve in 1X Extraction Buffer PTRBackground-subtracted data values (mean +/- SD) are graphed.
-
Interpolated concentrations of native MIF in mouse serum, plasma and cell culture supernatant samplesThe concentrations of MIF were measured in duplicates, interpolated from the MIF standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 0.5%, plasma (citrate) 2%, plasma (EDTA) 2%, plasma (heparin) 2%, and stimulated RAW 264.7 cell culture supernatant 0.3%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIF concentration was determined to be 1.06 µg/mL in serum, 286 ng/mL in plasma (citrate), 229 ng/mL in plasma (EDTA), 293 ng/mL in plasma (heparin), and 1.75 µg/mL in LPS-stimulated RAW 264.7 cell culture supernatant.
-
Interpolated concentrations of native MIF in RAW 264.7 cell extract and mouse spleen based on 0.49 µg/mL and 3.91 µg /mL extract loads, respectivelyThe concentrations of MIF were measured in duplicate and interpolated from the MIF standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MIF concentration was determined to be 9.8 µg/mL in RAW 264.7 cell extract and 1.1 µg/mL in mouse spleen extract.
Protocols
References
ab209885 has not yet been referenced specifically in any publications.
