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AngioKit™ - Human Angiogenesis Assay
ZHA-1000

AngioKit™ is supplied as growing cultures at the earliest stages of tubule formation in a 24 well plate format. It is designed so that test compounds, conditioned media or tissue explants can be added to the cultures within individual wells. The resulting effect on tubule formation can then be measured using TCS Cellworks AngioSys Image Analysis Software (ZHA-1800).

Control wells, which receive no treatment other than medium changes, form extensive networks of branching tubules over a period of 8-11 days. This allows both angiogenic inhibitors and angiogenic stimulators to be assessed. The necessary reagents for media changes are included in AngioKit™.

Positive and negative control compounds are available in the TCS Angiogenesis Control Reagent Kit (product code ZHA-1300). This kit contains AngioKit™-validated concentrated stock solutions of VEGF (2µg/ml positive controls) and Suramin (1mM negative controls) which require diluting in the medium supplied in the kit before use.

To run a complete AngioKit™ assay requires only a basic AngioKit™ (ZHA-1000), control reagents (ZHA-1300) and one of the available staining kits - see below for Related Products.

Kit Contents:

• 24 well pre-seeded tissue culture plate containing early stage co-cultures
• Extra sterile plate (use lid only)
• 5 x 25ml single use bottles of Optimised Growth Medium (one extra bottle to requirements is included for convenience)

For storage conditions and additional equipment required please read the protocol. Download the protocol here.

Example Tubule Formation with AngioKit™:

Fig 1a
Day 1 of tubule formation. Endothelial cells stained with antibody to von Willebrand factor (x100)

Fig 1b
Day 7 of tubule formation (x40)

Fig 1c
Day 14 of tubule formation (x40)

Tubules can also be visualised by staining for Collagen IV, a constitutive element of the extracellular matrix in neovascular tissue (Fig 2). The tubules produced also stain positive for the endothelial cell adhesion molecule PECAM-1 (Fig 3) and for ICAM-2 (not shown).

Fig 2
Tubules stained for collagen IV expression

Fig 3
Tubules stained for PECAM-1 expression

Molecules shown to be active in the TCS Cellworks AngioKit™ and V2a Kit™:

Note: that this represents only the molecules whose identity we know. We have tested  many  proprietary compounds from various laboratories that were active in both senses but which were supplied as coded samples whose structure is not known to us

Related Products

NEW ZHA-4000: V2a Kit™ - Vasculogenesis to Angiogenesis
This kit is supplied with cryopreserved co-culture cells, allowing users to begin the experiment at a time to suit them. Treatments can be added at any stage of the tubule development process to determine their effects, early during vasculogenesis or later for angiogenesis.

ZHA-1300: Angiogenesis Assay Control Kit
Contains VEGF and Suramin.

ZHA-1100: Angiogenesis Tubule Staining Kit ELISA
Contains Anti-CD31, secondary antibody and soluble substrate only.

ZHA-1225: Angiogenesis Tubule Staining Kit CD31
Contains Anti-CD31, secondary antibody and insoluble substrate.

ZHA-1125: Angiogenesis Tubule Staining Kit ELISA and CD31
Contains Anti-CD31, secondary antibody with both soluble and insoluble substrate.

ZHA-1250: Angiogenesis Tubule Staining Kit vWF
Contains Anti-von Willebrand Factor, secondary antibody and substrate.

ZHA-1200: Angiogenesis Tubule Staining Kit CD31 and vWF
Contains Anti-CD31, Anti-von Willebrand Factor secondary antibodies and substrates.

ZHA-1400: Optimised Growth Medium (100ml)

ZHA-1800: AngioSys Image Analysis software full license
A free 30 day use demonstration disc is available - conditions apply.

Anti-von Willebrand Factor polyclonal*

Anti-PECAM-1 (CD31) monoclonal*

Goat anti-mouse IgG; AP conjugate*

Donkey anti-sheep IgG; HRP conjugate*

* Bulk preparations of antibodies used in the various staining kits. Please contact TCS Cellworks for ordering information.