Technical datasheet
Description
The monoclonal antibody BV4 recognizes human beta3 integrin subunit present in Platelet glycoprotein GPIIb-IIIa (integrin alphaIIb/beta3, CD41/CD61) and in the vitronectin receptor (integrin alphaV/beta3, CD51/CD61). Intergins are a family of heterodimeric membrane glycoproteins expressed on diverse cell types which function as the major receptors for extracellular matrix and as cell-cell adhesion molecules. As adhesion molecules they play an important role in numerous biological processes such as platelet aggregation, inflammation, immune function, wound healing, tumour metastasis and tissue migration during embryogenesis. In addition integrins are involved in signaling pathways, transmitting signals both into an out from cells. All integrins consist of two non-covalently associated subunits, alpha and beta. At least 12 different alpha subunits and 8 beta subunits have been identified. The beta subunits all contain 56 conserved cysteines (except beta4 which has 48) which are arranged in four repeating units. The beta3 subunit is a 93kDa protein that contains a large loop in the N-terminus stabilized by intrachain disulphide bonding with the first cysteine-rich repeat.
Platelet glycoprotein GPIIb-IIIa is expressed on platelets and megakaryoblasts. It is constitutively expressed and becomes activated on triggered platelets. Platelet glycoprotein GPIIb-IIIa binds to fibrinogen, fibronectin, vWF, vitronectin and thrombospondin. Next to this it is also a receptor for several soluble adhesive proteins.Vitronectin receptor is expressed on endothelial cells, some B cells, monocytes/macrophages, platelets and tumour cells. Vitronectin receptor binds next to vitronectin to fibrinogen, vWF, thrombospondin, fibronectin, osteopontin and collagen. Defects in human beta3 integrin are a cause of Glanzmann thrombasthenia, which is an autosomal recessive disorder characterized by mucocutaneous bleeding and the inability of this integrin to recognize macromolecular or synthetic peptide ligands.
Cross Reactivity
| Cross reactant |
Reactivity |
| Bovine |
Yes |
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin and 0.02% sodium azide.
Species
Mouse IgG1
Application
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F2
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FC
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FS1
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IA
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IF
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IP1
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P3
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W4
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Yes
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No
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N.D.
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N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot
Application notes
IHC-F: Tissue sections were fixed in acetone. As negative control an irrelevant mouse IgG was used (Ref.2).
FS: Antibody clone BV4 inhibits the downstream activation, reducing the mitogenic effects of two of its ligands, VEGFR-2 and HIV-1-Tat. (Ref.1).
IP:clone BV4 precipitates beta3 integrin complexes from endothelial cell lysate.
IHC-P: Tissue sections were deparaffinized in xylene and pretreated with 10mmol/l sodium citrate buffer, pH 6.0
Use
For immunohistochemistry and Western blotting, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50. For functional studies, in vitro dilutions have to be optimized in user’s experimental setting.
Aliases
Platelet membrane glycoprotein IIIa, GPIIIa
Positive control
Endometrium
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least one year. The exact expiry date is indicated on the label.
References
1. Soldi, R et al; Role of alpha v beta 3 integrin in the activation of vascular endothelial growth factor receptor-2. EMBO J 1999, 18:882
2. Kimmins, S eta l: Immunohistochemical localization of integrin alpha v beta 3 and osteopontin suggests that they do not interact during embryo implantation in ruminants. Reprod Biol and Endocrinol. 2004, 2:19
3. Neto, D et al; Alpha-v-beta3 integrin expression in melanocytic nevi and cutaneous melanoma. J Cutan Pathol. 2007, 34: 851
4. Tang, N et al; N-terminal and C-terminal heparin-binding domain polypeptides derived from fibronectin reduce adhesion and invasion of liver cancer cells. BMC Cancer 2010, 10: 552
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.
References
1. Soldi, R et al; Role of alpha v beta 3 integrin in the activation of vascular endothelial growth factor receptor-2. EMBO J 1999, 18:882
2. Kimmins, S eta l: Immunohistochemical localization of integrin alpha v beta 3 and osteopontin suggests that they do not interact during embryo implantation in ruminants. Reprod Biol and Endocrinol. 2004, 2:19
3. Neto, D et al; Alpha-v-beta3 integrin expression in melanocytic nevi and cutaneous melanoma. J Cutan Pathol. 2007, 34: 851
4. Tang, N et al; N-terminal and C-terminal heparin-binding domain polypeptides derived from fibronectin reduce adhesion and invasion of liver cancer cells. BMC Cancer 2010, 10: 552