Mouse Predicted to work with:
Rat, Rabbit, Cow, Dog, Pig, Chinese hamster
Synthetic peptide corresponding to Mouse CTGF aa 250 to the C-terminus conjugated to keyhole limpet haemocyanin. Database link: P29268
This antibody gave a positive signal in CTGF Recombinant protein in Western Blot, as well as Mouse Lung FFPE tissue sections in IHC-P.
IF /ICC: HepG2 cell line.
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.
pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 38 kDa (predicted molecular weight: 38 kDa).
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 5 µg/ml.
Major connective tissue mitoattractant secreted by vascular endothelial cells. Promotes proliferation and differentiation of chondrocytes. Mediates heparin- and divalent cation-dependent cell adhesion in many cell types including fibroblasts, myofibroblasts, endothelial and epithelial cells. Enhances fibroblast growth factor-induced DNA synthesis.
Expressed in bone marrow and thymic cells. Also expressed one of two Wilms tumors tested.
ICC/IF image of ab125943 stained HepG2 cells. The cells were 100% methanol fixed (5 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 (ab125943, 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. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) HepG2 cells at 5µg/ml.
Western blot - Anti-CTGF antibody (ab125943)
Anti-CTGF antibody (ab125943) at 1 µg/ml + Connective Tissue Growth Factor (CTGF) Recombinant Protein at 0.1 µg
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 38 kDa Observed band size : 38 kDa Additional bands at : 29 kDa,33 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 1 minuteThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab125943 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
IHC image of CTGF staining in mouse lung formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. 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 ab125943, 5µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC 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.