ab32011 gave a positive result in rat and mouse brain tissue lysates. CAPS1 is known not to be expressed in heart (Speidel et al, J. Biol. Chem., Vol. 278, Issue 52, 52802-52809. PMID: 14530279). Mouse heart tissue lysate can be used as a negative control for ab32011.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: Use at a concentration of 1 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 146 kDa (predicted molecular weight: 146 kDa). Can be blocked with CAPS1 peptide (ab32785).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
CAPS1 (Ca2+-dependent activator protein for secretion) belongs to the CAPS/Cadps family which consists of two members, CAPS1 and CAPS2. The CAPS family proteins are involved in the secretion of different secretory substances in developing and postnatal brains. In addition CAPS1 regulates catecholamine release from neuroendocrine cells and is expressed predominantly in the brain.
Calcium-dependent activator protein for secretion 1 antibody
Western blot - CAPS1 antibody (ab32011)
All lanes : Anti-CAPS1 antibody (ab32011) at 1 µg/ml
Lane 1 : Brain (Rat) Tissue Lysate Lane 2 : Brain (Mouse) Tissue Lysate Lane 3 : Heart (Mouse) Tissue Lysate Lane 4 : Brain (Rat) Tissue Lysate with CAPS1 peptide (ab32785) at 1 µg/ml Lane 5 : Brain (Mouse) Tissue Lysate with CAPS1 peptide (ab32785) at 1 µg/ml Lane 6 : Heart (Mouse) Tissue Lysate with CAPS1 peptide (ab32785) at 1 µg/ml
Lysates/proteins at 10 µg per lane.
Secondary All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 146 kDa Observed band size: 146 kDa Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.
ab32011 is detecting a doublet at the expected molecular weight and both of these bands are blocked by the immunizing peptide. We are unsure what this doublet represents and would be interested in any feedback.
ICC/IF image of ab32011 stained human SH-SY5Y cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab32011, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Immunohistochemistry (PFA perfusion fixed frozen sections) - CAPS1 antibody (ab32011)This image is courtesy of an Abreview submitted by Dr Sophie Pezet
ab32011 staining CAPS1 in mouse brain tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue from 4% PFA perfused animals underwent overnight fixation in 4% paraformaldehyde, cryoprotected in 30% sucrose and cut using cryostat.The primary antibody was diluted, 1/300 (PBS + 0.3% Triton X100) and incubated with sample for 18 hours at 20°C. An abcam antibody ab60314, Chromeo 488 conjugated goat polyclonal to rabbit IgG, diluted 1/1000 was used as secondary.