This antibody gave a positive signal in the following Human Lysates:
TE 671 Whole Cell, HeLa Whole Cell - Hydroxyurea Treated (48hr, 2µM), Ramos Whole Cell, Placenta Tissue
ICC-IF: Hela cells untreated, Hela cells chloroquine treated.
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.
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
May play a role in ruffle formation and actin cytoskeleton organization. Seems to negatively regulate constitutive secretion.
Expressed in all tissues tested with highest expression in skeletal muscle and lowest in brain.
Cytoplasm > 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 staining CALCOCO2 in HeLa cells +/- Chloroquine (50μM, 24 hours). The cells were fixed with 4% paraformaldheyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab68588 at 5μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control, at 1/1000 dilution. The secondary antibodies were ab150081 (colored green) and ab150120 (pseudo-colored red) used at 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1 hour 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 All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
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
Kohler LJ et al. Effector Protein Cig2 Decreases Host Tolerance of Infection by Directing Constitutive Fusion of Autophagosomes with the Coxiella-Containing Vacuole. MBio7:N/A (2016).
Read more (PubMed: 27435465) »
Clough B et al. K63-Linked Ubiquitination Targets Toxoplasma gondii for Endo-lysosomal Destruction in IFN?-Stimulated Human Cells. PLoS Pathog12:e1006027 (2016).
Read more (PubMed: 27875583) »