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Catalog No: OKBB00331
Size:96 Tests
Price: $583.00
SKU
OKBB00331
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Datasheets/ManualsClick here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Product Info
Predicted Species ReactivityHuman
ApplicationELISA
ELISA Kit Detection MethodColorimetric, OD450 nm
ELISA Kit Duration~ 3 Hours
ELISA Kit PrincipleAviva's human MIF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for MIF has been precoated onto 96-well plates. Standards(E.coli, P2-A115) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for MIF is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human MIF amount of sample captured in plate.
ELISA Kit Range156 pg/ml - 10,000 pg/ml
ELISA Kit Reproducibility
Intra-Assay PrecisionInter-Assay Precision
Sample123123
Mean (pg/mL)144238266427162541776782
St. Dev.90.85206.6315121.9254.8393.4
%CV6.305.394.907.506.105.80
ELISA Kit Component
ComponentAmount
Lyophilized recombinant mouse C-MET standard10 ng/tube x 2
Anti-mouse C-MET Antibody Well Plate96 Wells
Sample diluent buffer30 mL
Biotinylated anti-mouse C-MET antibody130 uL, dilution 1:100
Antibody diluent buffer12 mL
Avidin-Biotin-Peroxidase Complex (ABC)130 uL, dilution 1:100
ABC diluent buffer12 mL
TMB color developing agent10 mL
TMB stop solution10 mL
Additional InformationRange: 156pg/ml-10,000pg/ml
::Principle: Aviva’s human MIF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human MIF specific-specific polyclonal antibodies were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human MIF amount of sample captured in plate.
::Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot experiment using standards and a small number of samples is recommended.
2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
4. Duplicate well assay is recommended for both standard and sample testing.
5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
6. Don’t reuse tips and tubes to avoid cross contamination.
7. To avoid to use the reagents from different batches together.
8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before using.
Background: Macrophage migration inhibitory factor (MIF) is a protein which in humans is encoded by the MIF gene.1, 2 This gene is located to human chromosome 22q11.2.3 It is remarkably small; it has 3 exons separated by introns of only 189 and 95 bp, and covers less than 1 kb.4 This gene encodes a lymphokine that may be involved in cell-mediated immunity, immunoregulation, and inflammation.5 MIF plays a role in the regulation of macrophage function in host defense through the suppression of anti-inflammatory effects of glucocorticoids. This lymphokine and the JAB1 protein might form a complex in the cytosol near the peripheral plasma membrane, which may possibly indicate a role in integrin signaling pathways. MIF also plays a central role in the toxic response to endotoxemia and possibly septic shock.Macrophage migration inhibitory factor has been reported to interact with COP9 constitutive photomorphogenic homolog subunit 5, CD74, BNIPL, and CXCR4. The standard product used in this kit is recombinant human MIF, consisting of 115 amino acids with the molecular mass of 12.5kDa.
Reconstitution and StorageStore at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
Sample Typecell culture supernates, serum, plasma (heparin, EDTA) and human milk
Sensitivity<20 pg/ml
Predicted Homology Based on Immunogen SequenceNo detectable cross-reactivity with any other cytokine.
Reference1. Weiser WY, Temple PA, Witek-Giannotti JS, Remold HG, Clark SC, David JR (October 1989). "Molecular cloning of
a cDNA encoding a human macrophage migration inhibitory factor". Proc. Natl. Acad. Sci. U.S.A. 86 (19): 7522–6.
2. Kozak CA, Adamson MC, Buckler CE, Segovia L, Paralkar V, Wistow G (June 1995). "Genomic cloning of mouse
MIF (macrophage inhibitory factor) and genetic mapping of the human and mouse expressed gene and nine mouse
pseudogenes". Genomics 27 (3): 405–11.
3. Budarf, M.; McDonald, T.; Sellinger, B.; Kozak, C.; Graham, C.; Wistow, G. : Localization of the human gene for
macrophage migration inhibitory factor (MIF) to chromosome 22q11.2. Genomics 39: 235-236, 1997.
4. Paralkar, V.; Wistow, G. : Cloning the human gene for macrophage migration inhibitory factor (MIF). Genomics 19:
48-51, 1994.
5. Leng L, Bucala R (February 2006). "Insight into the biology of macrophage migration inhibitory factor (MIF) revealed
by the cloning of its cell surface receptor.". Cell Research 16 (2): 162-8.
SpecificityNatural and recombinant human MIF
ImmunogenExpression system for standard: E.coli; Immunogen sequence: P2-A115
DescriptionFor quantitative detection of human MIF in cell culture supernatants, serum, plasma(heparin, EDTA) and human milk.
Gene SymbolMIF
Gene Full Namemacrophage migration inhibitory factor (glycosylation-inhibiting factor)
Alias SymbolsGIF, GLIF, Glycosylation inhibiting factor, L dopachrome isomerase, L dopachrome tautomerase, Macrophage migration inhibitory factor, Macrophage migration inhibitory factor (glycosylation inhibiting factor), MMIF, Phenylpyruvate tautomerase
NCBI Gene Id4282
Protein NameMacrophage migration inhibitory factor
Description of TargetMacrophage migration inhibitory factor (MIF) is a protein which in humans is encoded by the MIF gene. This gene is located to human chromosome 22q11.2. It is remarkably small; it has 3 exons separated by introns of only 189 and 95 bp, and covers less than 1 kb. This gene encodes a lymphokine that may be involved in cell-mediated immunity, immunoregulation, and inflammation. MIF plays a role in the regulation of macrophage function in host defense through the suppression of anti-inflammatory effects of glucocorticoids. This lymphokine and the JAB1 protein might form a complex in the cytosol near the peripheral plasma membrane, which may possibly indicate a role in integrin signaling pathways. MIF also plays a central role in the toxic response to endotoxemia and possibly septic shock.Macrophage migration inhibitory factor has been reported to interact with COP9 constitutive photomorphogenic homolog subunit 5, CD74, BNIPL, and CXCR4. The standard product used in this kit is recombinant human MIF, consisting of 115 amino acids with the molecular mass of 12.5kDa.
Uniprot IDP14174
Protein Accession #NP_002406.1
Nucleotide Accession #NM_002415.1
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