Immunohistochemical Analysis of Mouse Spleen

The following antibodies and reagents were utilized for immunohistochemical analysis of mouse spleen sections: PE-anti-CD4 (GK1.5); FITC-GL7 (RA3-6B2); APC-anti-IgD (all from BD Biosciences). Spleen cryostat sections (5–6 um) were prepared as previously described (15 (link)). Immunohistochemistry was performed as described (15 (link)) and stained sections were analyzed with a Leica DM4000 fluorescence microscope and Leica software (Leica Microsystems, Buffalo Grove, IL). The color intensity of the images was slightly enhanced using Adobe Photoshop CS4 (Adobe Systems, San Jose, CA) for better visualization and was carried out consistently between all sections while maintaining the integrity of the data.

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Wong E.B., Soni C., Chan A.Y., Domeier P.P., S.h.w.e.t.a.n.k., Abraham T., Limaye N., Khan T.N., Elias M.J., Chodisetti S.B., Wakeland E.K, & Rahman Z.S. (2015). B cell-intrinsic CD84 and Ly108 maintain germinal center B cell tolerance. Journal of immunology (Baltimore, Md. : 1950), 194(9), 4130-4143.

Publication 2015

Leica DM4000 fluorescence microscope

Anti igd Antibodies Buffalo Fitc Fluorescence microscope Immunohistochemistry Mouse Spleen

Corresponding Organization :

Other organizations : Pennsylvania State University, The University of Texas Southwestern Medical Center, UCLouvain

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Variable analysis

independent variables

Anti-CD4 (PE)
GL7 (FITC)
Anti-IgD (APC)

dependent variables

Immunohistochemical analysis of mouse spleen sections

control variables

Spleen cryostat sections (5–6 um)
Leica DM4000 fluorescence microscope
Leica software (Leica Microsystems, Buffalo Grove, IL)
Adobe Photoshop CS4 (Adobe Systems, San Jose, CA)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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