All lanes : Anti-CTHRC1 antibody (ab85739) at 1 µg/mlLane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lane 2 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431)Lane 3 : Blood Vessel: artery (Human) Membrane Lysate - adult normal tissue (ab28989)Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
Ab85739 staining CTHRC1 on E8.5 mouse embryo sections by immunohistochemistry. The tissue was paraformaldehyde fixed and permeabilized in PBST prior to blocking in NOVOCASTRA protein block for 30 minutes at 25°C. The primary antibody was diluted 1/100 and incubated with the sample for 48 hour at 4°C. An Alexa Fluor 555 conjugated donkey anti-rabbit antibody, diluted 1/1000, was used as the secondary. Nuclear staining shown with DAPI.See Abreview
ICC/IF image of ab85739 stained HepG2 cells. The cells were 4% PFA 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 (ab85739, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
All lanes : Anti-CTHRC1 antibody (ab85739) at 1 µg/mlLane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lane 2 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431)Lane 3 : Blood Vessel: artery (Human) Membrane Lysate - adult normal tissue (ab28989)Lane 4 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466) with Human CTHRC1 peptide (ab101727) at 1 µg/mlLane 5 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431) with Human CTHRC1 peptide (ab101727) at 1 µg/mlLane 6 : Blood Vessel: artery (Human) Membrane Lysate - adult normal tissue (ab28989) with Human CTHRC1 peptide (ab101727) at 1 µg/mlLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
IHC image of CTHRC1 staining in human skin melanoma formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab85739, 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. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.