Mouse IgM was prepared from normal serum by a multi-step process which includes delipidation, selective precipitation and tandem molecular sieve chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Mouse Serum and anti-Mouse IgM (µ chain specific). No reaction was observed against anti-Mouse IgG F(c). Some light chain cross-reactivity will occur with anti-Mouse IgG.
Formulierung:
Liquid (sterile filtered)
Application Verdünnung:
ELISA: user optimized, WB: user optimized
Anwendungsbeschreibung:
Biological Origin: Mouse. Application Notes: Mouse IgM whole molecule has been tested by SDS-Page and can be utilized as a control or standard reagent in SDS, Western Blotting, and ELISA experiments
Comparison of TI Ab responses in IUT, TBI, and control mice. Mice were immunized intraperitoneally with 25 µg of TNP-Ficoll. Sera were obtained 10 days after immunization and tested for IgM and IgG3 anti-TNP levels by sandwich ELISA. Each dot represents an individual mouse. Bar graphs represent average Ab levels SD. IUT mice were immunized starting at 12 weeks of age. TBI mice were challenged starting at 12 weeks after transplantation. Baseline total IgM and IgG serum levels were assessed as follows: 96-well ELISA plates were coated with goat anti-IgG (p/n orb347369) or goat anti-IgM Ab. Ig concentrations were calculated from standard curves generated with mouse IgM (p/n orb346283) and IgG standards.
IgM localizes within the conduit system of reactive LNs. (A) Confocal imaging of reactive and steady state auricular LNs from WT (C57BL/6J) or B cell-deficient (JHT) mice, 10 d after intradermal CFA immunization in the ears. Sections were stained for CD3, B220, collagen IV (col.IV), and IgM. Insets display high-magnification views of collagen IV+ conduits in the T cell zone. Bars, 100 µm (left panels), 10 µm (right panels). Data are representative of three experiments (two mice per condition and experiment). (B) Confocal imaging of auricular dLNs and contralateral ndLNs from Ccl19-Cre:RosatdT mice, 10 d after intradermal CFA injection in one ear. Sections were stained for collagen IV and IgM. tdT, tdTomato. Bars, 5 µm. Pictures are representative of three experiments (two mice per experiment). (C) Representative EM pictures of FRCs and conduits in steady state and auricular reactive LNs of WT and JHT mice, 10 d after intradermal CFA injection in one ear. Ultrathin sections were stained with 6-nm gold nanobeads specific for mouse IgM (highlighted with arrowheads), and the average number of beads located in the conduit lumen was evaluated for each condition. Bars, 2 µm (upper panels), 200 nm (lower panels). Data are representative of two experiments (one individual per condition and experiment). Results are expressed as mean SEM. *, P < 0.05, ****, P < 0.0001.
Lymph- and blood-borne soluble IgM does not access the conduit system. (A) Unimmunized JHT mice or animals that received an intradermal injection of CFA 10 d before in one ear were injected s.c. with fluorescent WGA (38 kD) with or without purified mouse IgM (1000 kD) and their auricular dLNs were harvested at the indicated time points. LN sections were stained for collagen IV (Col.IV) and IgM. Arrowheads indicate SCS. Insets show high-magnification views of the SCS and the underlying cortex. Bars, 100 µm (left panels), 20 µm (right panels). Data are representative of three experiments (two mice per condition and experiment). (B) JHT and WT mice were injected intradermally with CFA in one ear. 10 d later, JHT mice were supplemented with the serum of the immunized WT mice over a 24-h period and injected i.v. with fluorescent WGA 5 min before the harvest of reactive and contralateral auricular LNs. Data show confocal images from LN sections stained for collagen IV, PNAd, and IgM. Insets display high-magnification views of PNAd+ HEVs (yellow dashed line) and surrounding conduits. Bars, 200 µm (left panels), 50 µm (right panels). Data are representative of two experiments (three mice per experiment).
PLK1 regulates inflammatory cytokines induced by anti-CD40 and anti-IgM stimulation in vitro. Splenocytes were isolated from MLR.lpr mice, stimulated with (a) anti-CD40 antibody or (b) anti-IgM for 0.5 h and 24 h, with or without PLK1 blockade. Secretion of inflammatory cytokine in the culture supernatant were measured by ELISA, including IFNgamma, IL-1beta, TNFalpha, IL-6 and IL-10. The assays were performed in triplicate. Female MRL.lpr, 10-week-old, n = 3-6 per group. * indicates change between groups. indicates change within one group. *P < 0.05, **P < 0.01, P < 0.05, P < 0.01, P < 0.001. Immunoglobulin (Ig) standards, including mouse IgM, IgG or serum samples.
SDS-PAGE of
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