IgM Antibody [M15/8], Rabbit IgG

• Catalog Number: orb1672085
• Category: Antibodies
• Description: IgM Antibody [M15/8], Rabbit IgG
• Clonality: Recombinant
• Clone Number: M15/8
• Tested applications: Blocking, ELISA, FC, IHC-Fr, IP
• Reactivity: Human
• Isotype: IgG kappa
• Immunogen: BALB/c mice immunized with human IgM.
• Concentration: batch dependent
• Conjugation: Unconjugated
• UniProt ID: P01871
• Storage: Store at 4°C for up to 3 months. For longer storage, aliquot and store at -20°C.
• Buffer/Preservatives: PBS with 0.02% Proclin 300.
• Alternative names: mu-chain
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

ELISA of anti-IgM antibody on IgM Fc region peptide and a murine IgM antibody. Binding curves of the rmeric version of the anti-IgM antibody M15/8 (orb1672085; red line) and isotype control (anti-Fluorescein atnibody; 3.0; blue line) to an ELISA plate coated with either IgM Fc region peptide (Pr00108-15.5) at a concentration of 5 ug/ml (A) or murine IgM antibody (B; Anti-CD41 antibody; 1.0) again at 5 ug/ml. A 3-fold serial dilution from 10-000 to 0.17 ng/ml was performed using orb1672085. For signal detection- a 1:4000 dilution of HRP-lnti-r1 antibody was used. Both IgM Fc region peptide (monomeric ligand) and murine IgM antibody (multimeric ligand) were recognised by orb1672085.

Flow-cytometry using the anti-IgM M15/8. Daudi cells were stained with unimmunized r antibody (black line) or the rmeric version of M15/8 (orb1672085 - blue line) at a concentration of 10 ug/ml for 30 mins at RT. After washing- bound antibody was detected using anti-r JK (FITC-conjugate) antibody (129936) at 2 ug/ml and cells analyzed on a FACSCanto flow-cytometer.

Western Blot using anti-IgM antibody M15/8. Human spleen lysate (35 ug protein in RIPA buffer) was resolved on a 10% SDS PAGE gel and blots probed with the chimeric rsion of M15/8 (orb1672085) at 0.3 ug/ml before detection using an anti-rondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected band size for IgM is 180/900kDa for monomeric/pentameric IgM respectively. Here it is likely that the observed fragment is single glycosylated IgM heavy chain as s targetted to the Fc region of IgM (Cragg et al.- PMID: 10070880). orb1672085 successfully detected IgM in human spleen lysate.