The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 1µg for 106 cells.
Use at an assay dependent concentration. 30 - 60 mins at RT (ABC method). Formalin fixed paraffin embedded tissue sections require high temperature antigen unmasking with 10 mM citrate buffer, pH 6.0 prior to immunostaining.
Use at an assay dependent concentration.
Thought to provide a protective, lubricating barrier against particles and infectious agents at mucosal surfaces.
Expressed in corneal and conjunctival epithelia (at protein level). Overexpressed in ovarian carcinomas and ovarian low malignant potential (LMP) tumors as compared to the expression in normal ovarian tissue and ovarian adenomas.
Contains 2 ANK repeats. Contains 56 SEA domains.
Composed of three domains, a Ser-, Thr-rich N-terminal domain, a repeated domain containing more than 60 partially conserved tandem repeats of 156 amino acids each (AAs 12061-21862) and a C-terminal transmembrane contain domain with a short cytoplasmic tail.
Heavily O-glycosylated; expresses both type 1 and type 2 core glycans. Heavily N-glycosylated; expresses primarily high mannose and complex bisecting type N-linked glycans. May be phosphorylated. Phosphorylation of the intracellular C-terminal domain may induce proteolytic cleavage and the liberation of the extracellular domain into the extracellular space. May contain numerous disulfide bridges. Association of several molecules of the secreted form may occur through interchain disulfide bridges providing an extraordinarily large gel-like matrix in the extracellular space or in the lumen of secretory ducts.
Cell membrane. Secreted > extracellular space. May be liberated into the extracellular space following the phosphorylation of the intracellular C-terminus which induces the proteolytic cleavage and liberation of the extracellular domain.
Overlay histogram showing HeLa cells stained with ab697 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab697, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.