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You specify the PDGF BB and AA antibodies can be use for neutralization, could you send me a protocol please ?
Asked on Oct 31 2003
I have pasted the supplier's protocol below, but please do not hesitate to contact us should you have any other questions. Neutralization Assay for Anti-h-PDGF-BB using 3T3 cells Antibody: Anti-Human PDGF Procedure: 1) Resuspend 3T3 cells in regular culture media with 10% calf serum (Gibco lot# 1023609). Transfer 100ml culture/well to assay plate (5,000 cells/well). Incubate overnight. 2) Complete media change. Replace with 100ml assay media containing 0.5% calf serum. Incubate overnight. 3) Complete media change with assay media. 4) Serial dilute a-h-PDGF in assay media in triplicate. 5) Transfer assay media containing 8ng/ml h-PDGF to serial diluted antibody. Volume/well is 120 ml. Incubate 1 hour. 6) Transfer 100ml h-PDGF/anti-h-PDGFcomplex to cells in assay plate. Volume of each well is 200ml,containing 2 ng/ml h-PDGF and anti-h-PDGFas indicated below. Incubate 46 hours. 7) Add 20ml of Promega Substrate Cell Titer 96 Aqueous One Solution Reagent to each well. 8) Incubate 37oC and read at OD 490 nm.
Answered on Nov 04 2003