Human DISP1 Antibody

DISP1 in BG01V Human Embryonic Stem Cells.

DISP1 was detected in immersion fixed BG01V human embryonic stem cells using Sheep Anti-Human DISP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3549) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of DISP1 by Western Blot

Impaired Shh release from Disp−/− cells. (A) Alignment of targeted disp gene sequences from Disp−/− cells and from non-targeted (nt Ctrl) cells. (A′) Schematic representation of the Disp protein structure. An asterisk indicates the CRISPR/Cas9-generated stop codon introduced at position 323, deleting 11 of 12 TM domains that together represent ∼80% of the protein sequence. L1 and L2 indicate extracellular loops. TM2–TM6 (colored red) constitute the SSD. (B,C) Immunoblots of cellular (c) and released (into the medium, m) Shh (B) and unlipidated control C25SShhN (C) in nt Ctrl and Disp−/− cells in the presence of Scube2. Arrows indicate solubilized Shh and the arrowhead indicates accumulated cellular material in Disp−/− cells. (D) In the absence of Scube2, Shh processing into serum-free medium was abolished in nt Ctrl and Disp−/− cells. Instead, both cell types released similar amounts of unprocessed protein. In B,C,D, anti-beta -actin blots ( alpha beta -actin) and Ponceau S staining of residual serum albumin (PonS) serve as loading controls. (B′,C′,D′) Quantifications of relative Shh (B′,D′) and C25SShhN (C′) release from nt Ctrl and Disp−/− cells. Ratios of solubilized versus cellular Shh were determined and expressed relative to Shh release from nt Ctrl cells (black bars). Data are mean±s.d. n=21 in B′, n=8 in C′ and n=5 in D′. ****P<0.0001; ns, not significant (two-tailed unpaired t-test). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34308968), licensed under a CC-BY license. Not internally tested by R&D Systems.