This assay uses bisulfite-treated DNA and two site-specific probes for each marker, which bind to the associated methylated and unmethylated sequences.
We co-expressed a bright red-shifted firefly luciferase, along with the hNET and RQR8 - a small marker gene which binds the widely used antibody QBEnd/10 enabling flow cytometry and histochemistry.
To detect hypoxic regions, immunohistochemical analysis is performed with an anti‐HIF‐1α antibody and/or a hypoxic marker, pimonidazole, which binds to thiol‐containing proteins.
Similarly, staining for the +TIP marker EB1, which binds specifically to polymerising microtubules, shows that ephrin-A5/Fc treatment decreased the length of EB1 comets, indicating reduced microtubule polymerisation following ephrin-A5/Fc stimulation (Bieling et al., 2007).
Staining cells with cholera toxin B, a marker of membrane microdomains which binds to ganglioside GM1, revealed significantly reduced labeling in fibroblasts derived from the proband compared to both parental cell lines (Fig. 6), indicating that membrane lipid composition, and possibly formation of microdomains, are altered when CERS2 activity is reduced.
KI-67 is a marker for cell proliferation which binds a nuclear antigen and is thus a prime antibody in immunohistochemistry. Scoring methodologies derived from KI-67 immunostaining have shown promise as predictors of lethality in a range of cancers [ 1].
Here, it was applied fluorescent in situ hybridization labeling in suspension (FISHIS) [ 42] that uses an additional chromosome specific fluorescent marker which can quantitatively bind to chromosomes.
The chondrocyte hypertrophic marker COL10A1 is activated by MEF2C, which binds to conserved sequences in the promoter region of COL10A1 [ 38].
In combination with a specific primer which binds inside of the selectable marker (pyr4) and the primer which binds outside from the cassette we were able to very efficiently screen the mutant strains by PCR.
CTLA4 is a cell surface marker, associated with T-regulatory (T-reg) cell function, which binds to CD80 and CD86 on antigen presenting cells (APCs) sequestering them and preventing their interaction with CD28 on T-cells [ 101- 103].
