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V2a Kit™ - Vasculogenesis to Angiogenesis
ZHA-4000

The V2a Kit™ allows analysis of all aspects of the angiogenic pathway including; cell activation, cell proliferation, migration, elongation and anastomosis with complete flexibility of experimental timing. The kit contains a cryogenically preserved ampoule of cells so that you can seed the plate yourself and begin the assay at a time that suits you. V2a Kit™ is designed so that test compounds, conditioned media or tissue explants can be added to the culture at any time from the onset of vasculogenesis continuing through to advanced angiogenesis. The resulting effect on tubule formation can then be measured using TCS Cellworks AngioSys Image Analysis Software (ZHA-1800).

V2a Kit™ contains all of the cells and reagents necessary to complete a 24 well angiogenesis assay. The kit contains a cryogenically preserved ampoule of V2a cells, 24 well tissue culture plate, medium, supplements, control compounds and PECAM-1 staining reagents.

The advantage of this kit is that it can be run at a time to suit you, and that test compounds can be added as early as day 2 of the assay.

Control wells that receive no treatment other than medium changes form extensive networks of branching tubules over a period of one to two weeks. This allows both angiogenic inhibitors and angiogenic stimulators to be assessed. The necessary medium changes are included in the kit.

Positive and negative control compounds are included in V2a Kit™ consisting of validated concentrated stock solutions of VEGF (2ug/ml, positive control) and Suramin (1mM, negative control) which require diluting in the medium provided before use.

Kit Contents:

• Ampoule of cryopreserved V2a Cells
• 24 well tissue culture plate
• 96 well microtitre plate (for optional ELISA staining)
• 25ml Optimised Seeding Medium
• 125ml Optimised Growth Medium
• Seeding Medium Supplement
• Growth Medium Supplement
• Angiogenesis Control compounds (VEGF and Suramin)
• Mouse anti-human CD31 Primary Antibody
• Goat anti-mouse IgG Alkaline Phosphatase Secondary Antibody
• ρ-NPP soluble substrate and Buffer Tablets (for ELISA)
• BCIP/NBT insoluble substrate and Buffer Tablets (for permanent record of tubule staining)

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

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

Inhibitors	Enhancers
VEGF neutralising antibody	VEGF
VEGF antisense oligonucleotide	PIDGF
Collagen IV neutralising antibody	BFGF
PECAM-1 neutralising antibody	2-deoxyribose-1-P
VEGF-R antagonists	Most tumour cell conditioned media
Thrombospondin derivatives	Wound fluid
TNF alpha	Ascitic fluid from leiomyosarcoma patient
TGF b1	Ascorbic acid (biphasic response)
aVb3 integrin antagonists
Suramin
Curcumin
Fumagillin
TNP-740 (fumagillin analogue)
Colchicine
Cytochalasin B
Erucic acid
Ginsengosides
RGD peptide
Decorin
Endostatin
Thymine
Thalidomide and analogues
PAI-1 inhibitors MMP inhibitors
MGBG (methylglyoxalbis(guanylhydrozone)
Some conditioned media

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

ZHA-1000: AngioKit™ - Human Angiogenesis Assay
This kit is an in-vitro human angiogenesis model supplied as a pre seeded 24 well plate with co-culture cells. Untreated control wells form extensive tubule networks over a period of 6-11 days and allow for the testing of pro and anti-angiogenic compounds.

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.