The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/50 - 1/100.
Use a concentration of 10 µg/ml.
1/100. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
RelevanceCytokeratins are intermediate filament keratins found in the intracytoplasmic cytoskeleton of epithelial tissue. There are two types of Cytokeratins: the low weight, acidic type I cytokeratins and the high weight, basic or neutral type II. Cytokeratins are usually found in pairs comprising a type I Cytokeratin and a type II cytokeratin. The high molecular weight cytokeratins, which are the basic or neutral cytokeratins, comprise subtypes CK1 (67), CK2 (65.5), CK3 (64), CK4 (59), CK5 (58), CK6 (56), CK7 (54), CK8 (52.5) and CK9. The low molecular weight cytokeratins, which are the acidic cytokeratins, comprise subtypes CK10 (56.5), CK12 (56), CK13 (53), CK14 (50), CK16( 48), CK17 (46), CK18 (45), CK19(48) and CK20(46).
Overlay histogram showing A431 cells stained with ab86725 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab86725, 1/100 dilution) 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. Acquisition of >5,000 events was performed.
ICC/IF image of ab86725 stained DU145 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab86725 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Anti-HMW Cytokeratin antibody [34betaE12] (ab86725)
This product has been referenced in:
Kuramoto G et al. Preventive effect of oral mucosal epithelial cell sheets on intrauterine adhesions. Hum Reprod30:406-16 (2015).
Read more (PubMed: 25475585) »
Moinfar F et al. Use of keratin 35betaE12 as an adjunct in the diagnosis of mammary intraepithelial neoplasia-ductal type--benign and malignant intraductal proliferations. Am J Surg Pathol23:1048-58 (1999).
Read more (PubMed: 10478664) »