Immunohistochemical Analysis of Neural Markers

Tissue was collected and fixed, and sections were prepared as previously described [12 (link)]. The following antibodies were used: CC1 1: 250 (Millipore OP80), CNPase 1: 200 (Sternberger SMI91) (Covance, Princeton, NJ, USA), GFAP 1: 200 (Dako Z0334), Iba1 1: 150 (Wako 019-19741), MBP 1: 500 (Sternberger SMI99), Olig1 1: 600 (Chemicon AB5540) (EMD Millipore, Billerica, MA, USA), Olig2 1: 400 (Chemicon AB9610), O4 1: 250 (EMD Millipore MAB345), PDGFRA 1: 200 (Fitzgerald CD140a) (Fitzgerald, North Acton, MA, USA), pSMAD1/5/8 1: 50 (Cell Signaling 9511) (Cell Signaling Technology, Danvers, MA, USA), Alexa Fluor 488 goat anti-rabbit 1: 500, Alexa Fluor 594 goat anti-mouse IgG1 1: 500, Alexa Fluor 594 goat anti-mouse IgG2b 1: 500, Alexa Fluor 594 goat anti-mouse IgM 1: 500, and Alexa Fluor 647 goat anti-mouse IgM 1: 500 (Invitrogen) (Life Technologies, Grand Island, NY, USA). Confocal images were obtained using the Zeiss LSM 880 microscope at the Stanley Manne Children’s Research Institute.

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Dettman R.W., Birch D., Fernando A., Kessler J.A, & Dizon M.L. (2018). Targeted Knockdown of Bone Morphogenetic Protein Signaling within Neural Progenitors Protects the Brain and Improves Motor Function following Postnatal Hypoxia-Ischemia. Developmental neuroscience, 40(1), 23-38.

Publication 2018

GFAP pSMAD1/5/8 Alexa Fluor 488 goat anti-rabbit Alexa Fluor 594 goat anti-mouse IgG1 Alexa Fluor 594 goat anti-mouse IgG2b Alexa Fluor 594 goat anti-mouse IgM LSM 880 microscope

Alexa 594 Alexa fluor 488 Alexa fluor 647 Anti igm Antibodies Cell Children Cnpase Gfap Goat Igg1 Igg2b Mbp 1 Microscope Mouse Olig2 Rabbit Tissue

Corresponding Organization :

Other organizations : Neurology, Inc, Northwestern University

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Variable analysis

independent variables

Antibodies used for immunostaining: CC1, CNPase, GFAP, Iba1, MBP, Olig1, Olig2, O4, PDGFRA, pSMAD1/5/8

dependent variables

Immunostaining patterns and localization of the various markers

control variables

Tissue collection and preparation method as previously described [12]
Concentration and dilution of the antibodies used

controls

No positive or negative controls were explicitly mentioned in the provided information.

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