Mouse IgM Pincer^TM Assay

Catalog No: IM1009

This mouse IgM Pincer^TM Assay kit is to be used for the in vitro quantitative determination of mouse IgM concentrations in biological samples.

Immunoglobulin M (IgM) is a class of antibodies found in circulating body fluids. IgM molecule is usually a pentamer constructed from five basic monomer units which are covalently linked together with disulfide bonds¹. The presence of J chains also stabilizes the pentameric molecules². It has high molecular weight of 970,000 Daltons and 10 antigen binding sites. IgM can also exist as hexamer while the J chain is not presented in this form. IgM is secreted early on in an immune response and the half life of mouse IgM antibody has been reported as 1 day³ or less⁴ in vivo, whereas the half life in vitro was found to be greater than 6 days. It is resented on B cells and cannot cross the placental barrier. IgM promotes agglutination of red blood cells, phagocytosis and cell lysis.

This Mouse IgM pincer^TM Assay is a rapid, homogeneous antibody-based fluorescence assay for detecting and quantifying mouse IgM in biological samples such as ascitic fluids and hybridoma cell cultures. This assay consist two mouse IgM pincers that are the anti-mouse IgM specific antibody modified with two short complementary single-stranded DNAs. When mouse IgM is present, the two single-stranded DNAs associate to form a duplex and FRET (Fluorescence resonance energy transfer) is initiated between the two fluorophores located at the end of the two DNAs.  The amount of FRET signal is associated with the IgM concentration presented in the samples⁵. This assay is simple (one step) and fast (30 min).

Mouse IgM Pincer^TM Assay - Standard Curves

The following mouse IgM standard curve was prepared using the mouse IgM Pincer^TM Assay kit. This standard curve is for the purpose of illustration only, and should not be used to calculate unknowns. Each user should obtain his own data and standard curve.

Mouse IgM Pincer^TM Assay – Sensitivity And Specificity

Sensitivity: The minimum detectable concentration of mouse IgM by this assay is estimated to be 30 ng/ml.

Specificity: No cross reaction observed with human, rabbit, and goat IgG. There is some cross reaction (< 10%) observed with mouse IgG and the extent is depended on different IgG subtypies.

Reference:

1. Davis, A. C., and M. J. Shulman. (1989). IgM: molecular requirements for its assembly and function. Immunol. Today 10:118-122.
2. Koshland, M. E. (1985). The coming of age of the immunoglobulin J chain. Annu. Rev. Immunol. 3:425-453.
3. Britton, S., Wepsic, T. and MSller, G. (1968). Persistence of immunogenicity of two complex antigens retained in vivo. Immunology, 14: 491-501.
4. Fahey, J.L. & Sell, S. (1965). The immunoglobulins of mice. V. The metabolic properties of five im- munoglobulin class. J. Exp. Med., 122:41-58.
5. Heyduk, T., & Heyduk, E. (2005). Nucleic acid-based fluorescence sensors for detecting proteins. Anal. Chem., 77:1147-1156.

RESEARCH USE:

For research use only, not for use in diagnostic procedures.

Patent pending.

Product name	Measurements	Cat. #	Protocol	MSDS	Price	Order
Mouse IgM PincerTMAssay kit	100	MI1009-100			$120
Mouse IgM PincerTMAssay kit	384	MI1009-384			$280
384-well round-bottom low volume non-binding surface black polystyrene microplate	1 plate	163301	--		$9.50