The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
1/500 - 1/2000. Detects a band of approximately 45 kDa.
Use 1µg for 106 cells. ab170192-Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
FunctionAcid protease active in intracellular protein breakdown. Involved in the pathogenesis of several diseases such as breast cancer and possibly Alzheimer disease.
Tissue specificityExpressed in the aorta extrcellular space (at protein level).
Involvement in diseaseCeroid lipofuscinosis, neuronal, 10
Sequence similaritiesBelongs to the peptidase A1 family. Contains 1 peptidase A1 domain.
Post-translational modificationsN- and O-glycosylated.
Cellular localizationLysosome. Melanosome. Secreted, extracellular space. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. In aortic samples, detected as an extracellular protein loosely bound to the matrix (PubMed:20551380).
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Anti-Cathepsin D antibody [4G2] images
Flow Cytometry - Anti-Cathepsin D antibody [4G2] (ab40697)
Overlay histogram showing MCF7 cells stained with ab40697 (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 (ab40697, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Western blot - Cathepsin D antibody [4G2] (ab40697)
All lanes : Anti-Cathepsin D antibody [4G2] (ab40697) at 1/1000 dilution
Lane 1 : HepG2 cell lysate Lane 2 : Hep3B cell lysate Lane 3 : MDA-MB-231 cell lysate Lane 4 : MCF7 cell lysate
References for Anti-Cathepsin D antibody [4G2] (ab40697)
This product has been referenced in:
McNeill A et al. Ambroxol improves lysosomal biochemistry in glucocerebrosidase mutation-linked Parkinson disease cells. Brain137:1481-95 (2014).
Read more (PubMed: 24574503) »
Shen C et al. Global profiling of proteolytically modified proteins in human metastatic hepatocellular carcinoma cell lines reveals CAPN2 centered network. Proteomics12:1917-27 (2012).
Read more (PubMed: 22623320) »