Immunofluorescent analysis of Angiopoietin 2 in PC12 cells using an Angiopoietin 2 polyclonal antibody (Product # PA1-29640) at a dilution of 1:200 overnight at 4°C. Cells were fixed for 5 min in 100% methanol. Ag Retrieval/permeabilization: 1hr 1%BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween. Secondary detection was performed using a 488-conjugated Goat anti-rabbit secondary antibody at a dilution of 1:1000 (1 hr at room temperature). Ang2 staining is shown in yellow/green, plasma membranes in red and cell nuclei stained using DAPI (blue).
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||synthetic peptide amino acids 21-40 of Mouse Angiopoietin 2.|
|Storage buffer||whole serum diluted in 0.02M potassium phosphate, pH 7.2, with 0.15M NaCl|
|Contains||0.1% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||Assay-dependent|
|Western Blot (WB)||1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-29640 detects Angiopoietin 2 from human and mouse samples.
PA1-29640 has been successfully used in Western blot applications. There is reaction with serum in the cell supernatants, which results in strong background and makes it difficult to see the angiopoietins in cell supernatants. However when precipitated (using soluble Tie2) the signals are very good and strong.
The PA1-29640 immunogen is: Synthetic peptide: amino acids 21-40 of Mouse Angiopoietin 2.
Angiopoietin-1 (Ang-1) and Antiopoietin-2 (Ang2) are important for development of the endothelium, by regulating tyrosine phosphorylation of the membrane receptor Tie-2/Tek. Ang-1 binding to Tie-2/Tek causes phosphorylation of the receptor. Ang-2 competes for this binding, and thus blocks receptor phosphorylation. Ang-2 expression occurs at sites of vascular remodeling: dorsal aorta and major aortic branches, ovary, placenta and uterus. To our knowledge, this is the first time that an Ang antibody recognizes a band above 55 kD. This antibody (and the ang-1 antibody, GT28451) give a band at 75 kD which resembles the original size, because these are soluble highly glycosylated proteins, which should run higher that the calculated molecular weight of 55 kD.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.