Mfg Part Number
Ab01022-23.0
Vector Labs Ab01022-23.0 Anti-VEGF [G6-31], Rabbit IgG, kappa (200 μg)
Quick Overview
The anti-mVEGF Fab G6 underwent light chain randomisation by placing stop codons in positions 91-96 in CDR-L3 and an equimolar mix of oligonucleotides designed for mutagenesis was annealed to the mV401 stop template phagemid, followed by mutagenesis and E. coli electroporation. Binding selection was performed with mVEGF immobilised on immunoplates followed by solution-phase sorting with increasing stringency and single-point competetive ELISA to screen for high-affinity clones.
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| Product Class: | Purified |
|---|---|
| Clone ID: | G6-31 |
| Synonyms: | VEGF-A; Vascular endothelial growth factor; vascular permeability factor; VPF; MVCD1 |
| Amount: | 200 μg |
| Application Codes Clone: | ELISA; crystallisation; block; SPR; IF |
| Shipping Temperature: | Wet Ice |
| Buffer Composition: | PBS with 0.02% Proclin 300. |
| Original Format: | Fab |
| Storage Temperature: | Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at -20⁰C. |
| Chimeric Use Statement: | This chimeric rabbit antibody was made using the variable domain sequences of the original Human Fab format, for improved compatibility with existing reagents, assays and techniques. |
| Specificity Statement: | G6-31 recognises a VEGF epitope which is conserved between human and mouse VEGF and overlaps with the receptor binding surface. The KD of G6-31 binding to m-VEGF and h-VEGF is ~0.9 and 1.5 nM respectively. Alanine-scanning mutagenesis of solvent-accessible residues of h-VEGF identified 10 residues that result in >4-fold loss in binding affinity including F17, M18, Y21, Q22, Y25, I83,H86, Q89 and I91. |
| Application Notes: | A VEGF phage ELISA was used to measure binding affinity of G6-31 to individual alanine-substituted VEGF mutants versus wild type VEGF. Crystals of G6-31 and the receptor binding fragment of human VEGF have been solved at 3.6 Å (Fuh et al, 2005). Treatment of mice undergoing E2-induced PRLoma with G6-31 was used to block VEGF-A, causing reductions in the Ki-67-positive anterior cells, increases in TUNEL-positive anerior cells, and repair of the microvessel count by CD34-immunohistochemistry (Miyajima et al, 2010). Fluorescence quenching solution assay and SPR have been used to determine Kon and Koff at 25 degrees celsius (Lee et al, 2004). |
| Brand | Vector Labs |
|---|---|
| UNSPSC | 12161500 |