The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/50. PubMed: 19077055
Use at an assay dependent concentration. PMID 17724216
Use a concentration of 2 µg/ml.
Use at an assay dependent concentration. PubMed: 21490242
Use a concentration of 1 - 4 µg/ml. Detects a band of approximately 39 kDa (predicted molecular weight: 42 kDa).Can be blocked with CCN1 peptide (ab52125).
Promotes cell proliferation, chemotaxis, angiogenesis and cell adhesion. Appears to play a role in wound healing by up-regulating, in skin fibroblasts, the expression of a number of genes involved in angiogenesis, inflammation and matrix remodeling including VEGA-A, VEGA-C, MMP1, MMP3, TIMP1, uPA, PAI-1 and integrins alpha-3 and alpha-5. CYR61-mediated gene regulation is dependent on heparin-binding. Down-regulates the expression of alpha-1 and alpha-2 subunits of collagen type-1. Promotes cell adhesion and adhesive signaling through integrin alpha-6/beta-1, cell migration through integrin alpha-v/beta-5 and cell proliferation through integrin alpha-v/beta-3.
ICC/IF image of ab24448 stained HeLa 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 (ab24448, 1µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunocytochemistry/ Immunofluorescence - Anti-CCN1 antibody (ab24448)This image is courtesy of an anonymous abreview.
Immunocytochemistry/ Immunofluorescence analysis of human osteosarcoma cells labeling CCN1 with ab24448 at 1/100 dilution. Cells were fixed with paraformaldehyde and blocked with 0.05% triton-X100. The cells were blocked with 3% BSA for 1 hour at 20°C followed by incubation with ab24448 in TBS/Tween-20 + 1% BSA for 2 hours at 20°C. A Donkey anti-rabbit Cy3® secondary antibody was used at 1/200 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-CCN1 antibody (ab24448)This image is courtesy of an abreview submitted by Dr Torsten Lowin
Immunocytochemistry/ Immunofluorescence analysis of human synovial fibroblasts labeling CCN1 with ab24448 at 1/200 dilution. Cells were fixed with paraformaldehyde and permeabilized with methanol. The cells were blocked with 1% BSA for 1 hour at 25°C. The cells were incubated with PBS with ab24448 in 1% BSA for 2 hours at 25°C. A goat anti-rabbit Cy5® secondary antibody was used.
ab24448 staining Human myometrium. Staining can be observed in the cytoplasm and nuclear compartments. Left panel: with primary antibody at 2ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
Diamantopoulou Z et al. TIAM1 Antagonizes TAZ/YAP Both in the Destruction Complex in the Cytoplasm and in the Nucleus to Inhibit Invasion of Intestinal Epithelial Cells. Cancer Cell31:621-634.e6 (2017).
Read more (PubMed: 28416184) »
Zhang D et al. Fibulin-4 promotes osteosarcoma invasion and metastasis by inducing epithelial to mesenchymal transition via the PI3K/Akt/mTOR pathway. Int J Oncol50:1513-1530 (2017).
Read more (PubMed: 28339091) »