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You specify the PDGF BB and AA antibodies can be use for neutralization, could you send me a protocol please ? |
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ANSWER: |
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. |
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Currently thereis no stock for the PDGF. How long is the waiting time? |
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ANSWER: |
Unfortunately, we do not have it in stock. However, from placing your order it will take approximately 2 working days for us to obtain this antibody, followed by another 24 hrs to ship the product to you. |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
ab9704 staining PDGF BB in normal human skin tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections). Tissue underwent proteinase K mediated antigen retrieval. The primary antibody was used at 1 ug/ml and incubated with sample at 4°C overnight. A HRP-labeled polymer detection system was used with a non-alcohol soluble AEC chromogen.
Anti-PDGF BB antibody (ab9704) at 0.2 µg/ml + A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab52255) at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Observed band size : 18,29 kDa (why is the actual band size different from the predicted?)
Additional bands at : 45 kDa (possible dimer).
The 29kDa form is the propeptide.
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