Mouse MIP-2 ELISA Assay

$445.00

The Eagle Biosciences Mouse Macrophage Inflammatory Protein 2  (MIP-2) ELISA Assay Kit (enzyme-linked immunoassay kit) is intended for the quantitative determination of mouse Macrophage Inflammatory Protein 2  (MIP-2) concentrations in cell culture supernates, serum, and plasma. The Eagle Biosciences Mouse Macrophage Inflammatory Protein 2 (MIP-2) ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

Mouse MIP-2 ELISA Assay

The Mouse MIP-2 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 4 pg/mL
Dynamic Range: 7.8 – 250 pg/ml
Incubation Time: 3.5 hours
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Names: Macrophage Inflammatory Protein 2, CXCL2, C-X-C Motif Chemokine Ligand 2


Assay Background

Mouse macrophage inflammatory protein-2 (MIP-2), also known as CXCL2, was originally identified as a heparin-binding protein secreted by an LPS-stimulated mouse macrophage cell line. A cDNA clone encoding the protein was isolated from this cell line and characterized. Based on its protein and DNA sequences, mouse MIP-2 was classified as a member of the alpha (CXC) chemokine family of inflammatory and immunoregulatory cytokines.

Mouse MIP-2 cDNA encodes a 100 amino acid residue precursor protein from which the amino-terminal 27 amino acid residues are cleaved to generate the mature mouse MIP-2. The protein sequence of mouse MIP-2 shows approximately 63% identity to that of mouse KC, another mouse alpha chemokine. Mouse MIP-2 is also 60% identical to human GROβ and GROγ. Based on these protein sequence similarities, it is likely that mouse KC and MIP-2 are homologs of human GROα, β and γ chemokines. Since chemokines with protein sequence homology to human IL-8 have not been identified in mice, it has been suggested that the mouse KC and MIP-2 are functional homologs of human IL-8 in mice. A putative mouse homolog of the human IL-8 receptor beta (IL-8 Rβ) has also been cloned. This receptor shows 71% identity to human IL-8 Rβ and 68% identity to human IL-8 Rα. Both mouse KC and MIP-2 bind mouse IL-8 Rβ with high affinity.

Like human IL-8, mouse MIP-2 exhibits potent neutrophil chemotactic activity and may be a key mediator of neutrophil recruitment in response to tissue injury and infection. Increased MIP-2 expression has been found to be associated with neutrophil influx in various inflammatory conditions.


Related Products

Rat MIP-2 ELISA Assay Kit
Mouse GM-CSF ELISA Assay Kit
Mouse VEGF ELISA Assay

Additional Information

Assay Principle


The Eagle Biosciences Mouse Macrophage Inflammatory Protein 2 (MIP-2) ELISA Assay Kit employs quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for MIP-2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MIP-2 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for MIP-2 is added to the wells and binds to the combination of capture antibody-MIP-2 in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A colored product is formed in proportion to the amount of MIP-2 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven MIP-2 standard dilutions and MIP-2 sample concentration determined.

  1. Prepare all reagents and working standards as directed in the previous sections.
  2. Determine the number of microwell strips required to test the desired number of samples plus appropriate number of wells needed for running blanks and standards. Remove extra microwell strips from holder and store in foil bag with the desiccant provided at 2-8°C sealed tightly.
  3. Add 100 µL of Standard, control, or sample, per well, then add 50 µL of the working solution of Biotin-Conjugate to each well. Cover with the adhesive strip provided and incubate 2 hours at RT. Adequate mixing is very important for good result. Use a mini-vortexer at the lowest frequency.
  4. Aspirate each well and wash, repeating the process three times for a total of four washes. Wash by filling each well with Wash Buffer (350 µL) using a squirt bottle, manifold dispenser or auto-washer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
  5. Add 100 µL of the working solution of Streptavidin-HRP to each well. Cover with a new adhesive strip and incubate for 30 minutes at RT Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash.
  7. Add 100 µL of Substrate Solution to each well. Incubate for 10-20 minutes at RT. Avoid placing the plate in direct light.
  8. Add 100 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm.(optionally 630nm as the reference wave length;610-650nm is acceptable)

Documents

Product Manual


 

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

Publications

References


  1. Wolpe, S.D. et al. (1989) Proc. Natl. Acad. Sci. USA 86:612.
  2. Tekamp-Olson, P. et al. (1990) J. Exp. Med. 172:911.
  3. Schall, T. (1994) in The Cytokine Handbook, 2nd ed., A. Thomson, ed., Academic Press, New York, p. 419.
  4. Driscoll, K.E. (1994) Exp. Lung Res. 20:473.
  5. Heinrich, J. and R. Bravo (1995) J. Biol. Chem. 270:4987.
  6. Standiford, T.J. et al. (1996) J. Leukoc. Biol. 59:24.
  7. Su, Y.H. et al. (1996) J. Virol. 70:1277.
  8. Greenberger, M.J. et al. (1996) J. Infect. Dis. 173:159.
  9. Seebach, J. et al. (1995) J. Immunol. 155:4367.
  10. Standiford, T.J. et al. (1995) J. Immuno. 155:2222

Product Citations