This antibody gave a positive signal in HepG2 whole cell lysate as well as the following tissue lysates: Human Heart; Human Spleen; Human Bone Tumour; Human Bone Marrow.
This antibody gave a positive result in IF in the following formaldehyde fixed cell lines: DU145.
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Protein Z, vitamin K dependent plasma glycoprotein antibody
Vitamin K-dependent protein Z antibody
Western blot - Anti-Protein Z antibody (ab85317)
All lanes : Anti-Protein Z antibody (ab85317) at 1 µg/ml (Milk Blocking - 5%)
Lane 1 : Human heart tissue lysate - total protein (ab29431) Lane 2 : Human spleen tissue lysate - total protein (ab29699) Lane 3 : Human bone tumor tissue lysate - total protein (ab29359) Lane 4 : Bone Marrow (Human) Tissue Lysate - adult normal tissue Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
The band observed at 42 kDa could potentially be a cleaved form of Protein Z due to the presemce of a 23 amino acid signal peptide.
Immunocytochemistry/ Immunofluorescence - Anti-Protein Z antibody (ab85317)
ICC/IF image of ab85317 stained DU145 cells. The cells were 4% formaldehyde fixed (10 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 ab85317 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.