Sandwich ELISA analysis of IgM binding using Mouse anti Human IgM antibody, clone M15/8 (MCA1662) as a capture reagent and biotinylated Goat anti Human IgM (AHP1764) as a detection regent with purified human IgM (5275-5504) as antigen for the generation of the standard curve. Subsequent detection is by HRP conjugated streptavidin and substrate. Microtitre plate is read at O.D. 450 nm on the Bio-Rad iMark Microplate Absorbance Reader (Bio-Rad Catalog 168-1135). Serum samples at 1:10k (green) and 1:100k (blue), similarly plasma samples 1:10k (orange) and 1:100k (red) dilutions are shown
Native Human IgM is purified from human plasma and demonstrated a single band in electrophoresis. Native Human IgM has been used as the IgM antigen in the development of a sensitive ELISA assay for the measurement of IgM in human samples.
Purified immunoglobulin from human plasma - lyophilised
Reconstitute with 1ml 250mM Sodium Chloride. Further dilutions may then be made in sterile Phosphate Buffered Saline. Care should be taken during reconstitution as the protein may appear as a film at the bottom of the vial. Bio-Rad recommend that the vial is gently mixed after reconstitution.
Purified IgM prepared by delipidation, salt fractionation and ion-exchange chromatography.
>95% Single band in high resolution protein electrophoresis on Corning HRPE agarose gel. 0.5 µl of IgM solution at 50 mg/ml in isotonic saline.
Approx. Protein Concentrations
5.0 mg/ml after reconstitution
Reagents In The Kit
Preparing The Antibody
Phosphate buffered saline.
Store at -20oC only. Storage in frost-free freezers is not recommended. This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the protein. Should this product contain a precipitate we recommend microcentrifugation before use.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.