Fourteen-week-old mice were immunized subcutaneously with 150 μg of myelin oligodendrocyte glycoprotein peptide (residues 35–55; MOG35–55; Bio-Synthesis, TX) emulsified in complete Freund’s adjuvant (CFA; Chondrex, Redmond, WA) containing 2.5 mg/ml heat-killed Mycobacterium tuberculosis. Mice were injected intraperitoneally with 250 ng Bordetella pertussis toxin (Chondrex) the day of, and again, 2 days following MOG35–55 immunization. At 21–28 days post-immunization, mice were euthanized for histological analyses. The mice were euthanized by CO2 asphyxiation for 10 min from a compressed gas tank followed by decapitation or cardiac perfusion. Following perfusion, mice were immediately decapitated. EAE clinical manifestations of progressive ascending paralysis were assessed daily using a standard scoring system: 0 = no overt symptoms, 1= loss of tail tone, 2 = hind limb paresis, 3 = complete hind limb paralysis, 4 = hind limb paralysis and forelimb paresis, and 5 = moribund or dead. The mice were weighed daily to ensure weight loss did not exceed 25% of starting weight.
Mog35 55
Manufactured by Chondrex
MOG35–55 is a laboratory product offered by Chondrex. It is a synthetic peptide corresponding to amino acids 35–55 of myelin oligodendrocyte glycoprotein (MOG). This product is intended for research and scientific applications.
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3 protocols using mog35 55
1
Induction of Experimental Autoimmune Encephalomyelitis
2
Induction of Experimental Autoimmune Encephalomyelitis in Mice
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Eight- to nine-week-old mice were immunized subcutaneously with 150 μg myelin oligodendrocyte glycoprotein (MOG35–55; Bio-Synthesis, City, TX) emulsified in complete Freund’s adjuvant (CFA; Difco Laboratories, Detroit, MI) containing 2.5 mg/ml heat-killed Myobacterium tuberculosis. Mice were injected intraperitoneally with 250 ng Bordetella pertussis toxin (Chondrex, Redmond, WA) the day of, and 2 days following, MOG35–55 immunization. At 21 days post-immunization, the mice were euthanized for the histological analyses. EAE clinical manifestations of progressive ascending paralysis were assessed daily using a standard scoring system: 1 = loss of tail tone, 2 = hind limb paresis, 3 = complete hind limb paralysis, 4 = hind limb paralysis and forelimb paresis, and 5 = moribund or dead. Mice were weighed daily to ensure weight loss did not exceed 25% of starting weight. All EAE data were gathered in two independent experiments from cohorts of the same 12 WT mice and ten KO mice.
3
Induction of Experimental Autoimmune Encephalomyelitis
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EAE was actively induced in mice using MOG35-55(Sigma-Aldrich) as previously described (5 (link)). In brief, MOG35-55 was diluted to 10 mg/mL in 0.9% physiological saline and combined with an equivalent volume of complete Freund's adjuvant (CFA, Chondrex) and tuberculin H37Ra (DIFCO) at a concentration of 4 mg/mL. The solution was pumped into an oil-in-water form by syringe to prepare an induction antigen emulsion. Mice were subcutaneously injected with 0.1 mL of the emulsifier at 4 points on both sides of the spine and then intraperitoneally injected with 0.5 mL of pertussis toxin (PT, American LBL Company) twice at 0 and 48 h post-immunization. One week later, the mice received a booster injection of the MOG35-55 solution.
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