The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 2.5 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).Can be blocked with Caspase-14 peptide (ab72738).
Use a concentration of 5 µg/ml.
Believed to be a non-apoptotic caspase which is involved in epidermal differentiation. Seems to play a role in keratinocyte differentiation and cornification. Probably regulates maturation of the epidermis by proteolytically processing filaggrin.
Expressed in keratinocytes of adult skin suprabasal layers (from spinous layers to the stratum granulosum and stratum corneum) (at protein level). Expressed in keratinocytes of hair shaft and sebaceous glands (at protein level). In psoriatic skin only expressed at very low levels.
ab45415 stained HepG2 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 ab45415 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 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. This antibody also gave a positive result in 100% methanol fixed (5min) HeLa cells at 5ug/ml.
Western blot - Anti-Caspase-14 antibody (ab45415)
All lanes : Anti-Caspase-14 antibody (ab45415) at 1 µg/ml
Lane 1 : Jurkat cell lysate
Lane 2 : Jurkat cell lysate + blocking peptide
Secondary anti-rabbit IgG
Predicted band size : 27 kDa Observed band size : 27 kDa
Asselin-Labat ML et al. Gata-3 negatively regulates the tumor-initiating capacity of mammary luminal progenitor cells and targets the putative tumor suppressor caspase-14. Mol Cell Biol31:4609-22 (2011).
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