The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 5 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 52 kDa).
Use at an assay dependent concentration. PubMed: 23209807
FunctionMay play a role in ruffle formation and actin cytoskeleton organization. Seems to negatively regulate constitutive secretion.
Tissue specificityExpressed in all tissues tested with highest expression in skeletal muscle and lowest in brain.
Cellular localizationCytoplasm > perinuclear region. Golgi apparatus. Cytoplasm > cytoskeleton. According to PubMed:7540613, localizes to nuclear dots. According to PubMed:9230084 and PubMed:12869526, it is not a nuclear dot-associated protein but localizes predominantly in the cytoplasm with a coarse-grained distribution preferentially close to the nucleus.
ab68588 stained HeLa cells. The cells were 4% PFA fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab68588 at 1µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Western blot - CALCOCO2 antibody (ab68588)
All lanes : Anti-CALCOCO2 antibody (ab68588) at 1 µg/ml
Lane 1 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate Lane 2 : HeLa Whole Cell Lysate - Hydroxyurea Treated (48hr, 2µM) Lane 3 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate Lane 4 : Human placenta tissue lysate - total protein (ab29745)
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution developed using the ECL technique
IHC image of CALCOCO2 staining in Human Cerebral Cortex FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab68588, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
References for Anti-CALCOCO2 antibody (ab68588)
This product has been referenced in:
Jo C et al. Nrf2 reduces levels of phosphorylated tau protein by inducing autophagy adaptor protein NDP52. Nat Commun5:3496 (2014).
Read more (PubMed: 24667209) »
Baxt LA & Goldberg MB Host and bacterial proteins that repress recruitment of LC3 to Shigella early during infection. PLoS One9:e94653 (2014).
Read more (PubMed: 24722587) »