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Your Position: Home > All Other Proteins > Integrin alpha V beta 5 > Biotinylated Human Integrin alpha V beta 5 (ITGAV&ITGB5) Heterodimer Protein, His,Avitag&Tag Free

Biotinylated Human Integrin alpha V beta 5 (ITGAV&ITGB5) Heterodimer Protein, His,Avitag™&Tag Free

INTRODUCTION

QC & Assay Protocol

Citation & Background

Synonym
Integrin alpha V beta 5,ITGAV&ITGB5
Source
Biotinylated Human ITGAV&ITGB5 Heterodimer Protein, His,Avitag&Tag Free (IT5-H82Wa) is expressed from human 293 cells (HEK293). It contains AA Phe 31 - Val 992 (ITGAV) & Gly 24 - Asn 719 (ITGB5) (Accession # NP_002201.1 (ITGAV) & P18084-1 (ITGB5)).
Predicted N-terminus: Phe 31 (ITGAV) & Gly 24 (ITGB5)
Request for sequence
Molecular Characterization
Biotinylated Human ITGAV&ITGB5 Heterodimer Protein, His,Avitag&Tag Free, produced by co-expression of ITGAV and ITGB5, has a calculated MW of 114.7 kDa (ITGAV) and 81.9 kDa (ITGB5). Subunit ITGAV is fused with an acidic tail at the C-terminus and followed by a polyhistidine tag and an Avi tag (Avitag™) and subunit ITGB5 contains no tag but a basic tail at the C-terminus. The non-reducing (NR) protein migrates as 140-150 kDa (ITGAV) and 90-98 kDa (ITGB5) respectively due to glycosylation.
Biotinylation
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
Biotin:Protein Ratio
The biotin to protein ratio is 0.5-1 as determined by the HABA assay.

Endotoxin
Less than 1.0 EU per μg by the LAL method.
Purity
>95% as determined by SDS-PAGE.
Formulation
Lyophilized from 0.22 μm filtered solution in 50 mM Tris, 150 mM NaCl, pH7.5. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
No activity loss was observed after storage at:
1. 4-8^°C for 12 months in lyophilized state;
2. -70^°C for 3 months under sterile conditions after reconstitution.

SDS-PAGE

Biotinylated Human ITGAV&ITGB5 Heterodimer Protein, His,Avitag&Tag Free on SDS-PAGE under non-reducing (NR) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.

Bioactivity-ELISA

Biotinylated Human ITGAV&ITGB5 Heterodimer Protein, His,Avitag&Tag FreeBiotinylated Human ITGAV&ITGB5 Heterodimer Protein, His,Avitag&Tag Free (Cat. No. IT5-H82Wa) ELISA bioactivity

Immobilized Human Vitronectin at 5 μg/mL (100 μL/well) can bind Biotinylated Human ITGAV&ITGB5 Heterodimer Protein, His,Avitag&Tag Free (Cat. No. IT5-H82Wa) with a linear range of 0.01-0.156 μg/mL (QC tested).

Protocol

Immobilized Human Fibronectin at 5 μg/mL (100 μL/well) can bind Biotinylated Human ITGAV&ITGB5 Heterodimer Protein, His,Avitag&Tag Free (Cat. No. IT5-H82Wa) with a linear range of 0.156-5 μg/mL (Routinely tested).

Protocol

Background
Integrin alpha V beta 5 (ITGAV & ITGB5) is expressed on a wide variety of cell types including keratinocytes, fibroblasts, adhesive monocytes, embryonic stem cells, and select endothelium and epithelium. ITGAV & ITGB5 binds ligands containing an RGD motif, notably vitronectin. Growth factors that increase PKC activity, such as VEGF or TGF alpha, promote ITGAV & ITGB5-mediated angiogenesis while alpha V beta 3, which may be expressed in the same cell, responds to FGF-basic and TNF alpha. An inhibitor of both down regulates tumor angiogenesis. During lung inflammation, up regulation of ITGAV & ITGB5 on myofibroblasts or infiltrating lymphocytes may contribute to fibrosis by freeing TGF beta from latency.
References
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.