Technical Data
Angiostatin Kringles 1-4
Embryonic vascular system undergoes a series of complex, highly regulated series of events involving differentiation, migration and association of primitive endothelial cells. This process is termed vasculogenesis. A further remodeling of the primitive vascular system forms the mature cardiovascular system. This process is known as angiogenesis (sprouting of new capillary vessels from pre-existing vasculature). Angiogenesis accounts for the formation of vasculature into previously avascular organs such as brain and kidney. Angiogenic activity in the adult is required during the normal tissue repair, and for the remodeling of the female reproductive organs (ovulation and placental development). Certain pathological conditions, such as tumor growth and diabetic retinopathy, also require angiogenesis.

Recent studies have identified several proteolytic fragments or cryptic domains of proteins that act as inhibitors of angiogenesis. These include fragments of plasminogen such as Angiostatin protein (kringles 1-4) and kringles 1-5, C-terminal proteolytic fragment of Collagen XVIII (Endostatin protein), the NC10 domain of collagen 15 (Restin), the C-terminal hemopexin-like domain of MMP-2 (PEX), the N-terminal fragment of prolactin, and the N-terminally truncated platelet factor. Angiostatin protein, a proteolytic fragment of plasminogen, is comprised of the first four kringle regions. It prevents the growth of endothelial cells, and its systemic administration inhibits the growth of primary carcinomas in mice. Kringles 1-3 fragment has a greater inhibitory activity than the kringles 1-4 fragment. The protease-activated kringles 1-5 is the most potent plasminogen fragment with over 50-fold greater endothelial cell specific inhibitory activity. Its systemic administration inhibited the growth of fibrosarcoma and significantly reduced neovascularization.

Suitable for use in ELISA, Western Blotting. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000-1:5000 using chemiluminescence. Human angiostatin (kringles 1-4) is ~50kD.

ELISA: 1:10,000-1:100,000; using 50-100ng of control peptide/well

Immunohistochemistry & Immunofluorescence: Not tested. We recommend the use of affinity purified antibody at 2-20ug/ml in formaldehyde fixed tissue.

Optimal dilutions to be determined by the researcher.

Positive Control: A2294-35B
Human angiostatin protein formulated in SDS-PAGE sample buffer (reduced). This preparation is biologically inactive. It is not suitable for ELISA or other applications where native protein is required.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
100ul-20CBlue IceHumanRabbit
As reported
Angiostatin protein (~50kD; kringles 1-4) is prepared form individual that have been negative for HBsAg, HIV, and HCV. It is 95% pure as determined by SDS-PAGE. Purified protein is free from plasmin or plasminogen.
Neat serum
Supplied as a lyophilized powder from PBS, 0.05% sodium azide.
Recognizes human Angiostatin.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
General References: Peterson Te et al (1990) JBC 265, 6104-6111; Forsgren m et al (1987) FEBS Lett. 213, 254-260; Malinowski DP et al (1984) Biochemistry 23, 4243-4250; O'Reilly MS et al (1994) Cell 79, 315-328; Sim BK et al (1997) Cancer Res. 57, 1329-1334; Wu Z et al (1997) BBRC 236, 651. O'Reilly MS et al (1994) Cell 79, 315-328, Cao E et al (1999) PNAS 96, 5728; Cao Y et al (1997) JBC, 272, 22924; Peterson Te et al (1990) JBC 265, 6104-6111; Forsgren m et al (1987) FEBS Lett. 213, 254-260; Malinowski DP et al (1984) Biochemistry 23, 4243-4250; ; Sim BK et al (1997) Cancer Res. 57, 1329-1334; Wu Z et al (1997) BBRC 236, 651.