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Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Rat Hippocalcin.
(Peptide available as ab25848.)
Our Abpromise guarantee covers the use of ab24560 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).|
|IHC-P||1/350. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IHC (PFA fixed)||Use a concentration of 0.2 µg/ml.|
|ICC/IF||Use a concentration of 5 µg/ml.|
|IHC-Fr||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
Immuofluorescent staining for Hippocalcin in the rat hippocampus using Rabbit polyclonal to Hippocalcin (ab24560). This figure is a montage of pictures acquired with a X10 objective and shows expected abundance of staining in parts of the hippocampus such as the CA1 and CA3 and an absence of staining (as expected) in the cortex and the corpus callosum. ab24560 was used at 1/1000 (0.2µg/ml) incubated overnight at room temperature. Secondary antibody used was anti-rabbit Alexa Fluor 488 at 1/1000 incubated for 2 hours at room temperature. Rat brain tissue was perfusion fixed with 4% PFA followed by overnight post-fixation in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. 30µm coronal sections were cut on a cyrostat and immunohistochemistry performed by the 'free floating' technique.
ICC/IF image of ab24560 stained SKNSH 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 ab24560 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.
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