Skip to main content
. 2022 May 25;92(2):279–291. doi: 10.1002/ana.26380

FIGURE 5.

FIGURE 5

Detection of ZSCAN1 autoantibodies with slot‐blot Western blotting using ROHHAD CSF. Whole cell‐lysates from HEK293T cells expressing transfected with full‐length human ZSCAN1 cDNA were separated on a 1‐well 4–12% Tris–HCl protein gel, transferred to a PVDF membrane and immunoblotted in a slot‐blot device (BioRad). Primary and secondary antibodies were added to lanes as indicated. Primary antibodies were loaded in lanes 1–12. Lanes 13 and 15 served as secondary‐only controls. Primary antibodies are as follows: lanes 1 through 7: ROHHAD 1 through 7 CSF (1:200); lanes 8 through 12: controls 1–5 CSF (1:200); lane 13: blank; lane 14: commercial antibody to ZSCAN1 (Sigma, rabbit, 1:2000); lane 15: blank, lane 16: commercial antibody to Flag (CST, rabbit, 1:2000). Secondary antibodies to visualize human IgG loaded in lanes 1–12: goat anti‐human IgG (LICOR680). Secondary antibodies to visualize commercial antibodies to ZSCAN1 and FLAG lanes 14 and 16: goat anti‐rabbit IgG (LICOR800).