Spag6 Knockout Immune Response

C57/SV129 WT were irradiated and reconstituted i.v with 5 million bone marrow cells from WT or Spag6KO as previously described40 (link)41 (link). After 6 weeks, mice were footpad and i.p immunized with 10 μg 4-hydroxy-3-nitrophenylacetyl coupled to keyhole limpet hemocyanin at a ratio of 27:1 (Bioresearch Technologies) in 4 mg alum. Mice were bled at day 7 and organs harvested at day 14. NP-specific IgM and IgG1 was determined by ELISA using NP-14–BSA (15 μg/ml; Biosearch Technologies) as previously described42 (link).

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Cooley L.F., El Shikh M.E., Li W., Keim R.C., Zhang Z., Strauss J.F., Zhang Z, & Conrad D.H. (2016). Impaired immunological synapse in sperm associated antigen 6 (SPAG6) deficient mice. Scientific Reports, 6, 25840.

Publication 2016

NP-14–BSA

Alum Bone marrow cells Elisa Igg1 Keyhole limpet hemocyanin Mice

Corresponding Organization : Virginia Commonwealth University

Other organizations : Queen Mary University of London, William Harvey Research Institute, Huazhong University of Science and Technology

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Variable analysis

independent variables

Reconstitution of irradiated mice with WT or Spag6KO bone marrow cells

dependent variables

NP-specific IgM and IgG1 levels measured by ELISA

control variables

C57/SV129 WT mice
Footpad and i.p immunization with 10 μg 4-hydroxy-3-nitrophenylacetyl coupled to keyhole limpet hemocyanin in 4 mg alum
Time points: Day 7 for blood collection, Day 14 for organ harvest

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