Anti-Nucleolin抗体[3G4B2] (ab14966)

I have been informed that the statement regarding IP for this antibody (which is the same as clone 3G4B2) comes from the reference Gotzmann J et al. Two-dimensional electrophoresis reveals a nuclear matrix-associated nucleolin complex of basic isoelectric point. Electrophoresis 18:2645-53 (1997). PubMed: 9527495 I found the article in PubMed, but the full length article was not available so I cannot quote a dilution value for IP, I'm very sorry I cannot help you more. I hope you will be able to access the paper and find this information, my apologies for the delay,

Thank you for your enquiry and patience. I was able to obtain the following information from the source of this antibody. Human nucleolin is consistently described as having two isoforms, with molecular weights of 95 kDa and 90 kDa. Ab14966 recognizes these isoforms and on a Western blot, there is a strong band at 95 kDa and a much fainter band at 90 kDa. This image will be added to our online datasheet. Here is the Western blot protocol as well: 1. Perform SDS-PAGE on a cell lysate sample (cell lysis buffer: 50mM Tris-HCl, pH7.4; 1% NP-40; 0.25% sodium deoxycholate; 150mM NaCl; 1mM EGTA; 1mM PMSF; 1ug/ml each aprotinin, leupeptin, pepstatin; 1mM Na3VO4; 1mM NaF) and transfer the proteins to nitrocellulose. Wash the blotted nitrocellulose twice with water. 2. Block the blotted nitrocellulose in freshly prepared PBS containing 3% nonfat dry milk (PBS-MLK) for 60 minutes at RT 3. Incubate the nitrocellulose with a 1:1000-1:5000 dilution of anti-Nucleolin diluted in freshly prepared PBS-MLK overnight with agitation at 4°C. 4. Wash the nitrocellulose twice with water. 5. Incubate the nitrocellulose in the secondary reagent of choice (a goat anti-mouse HRP conjugated IgG, 1:5000 dilution was used) in PBS-MLK for 1.0 hours at RT with agitation. 6. Wash the nitrocellulose with water twice. 7. Wash the nitrocellulose in PBS-0.05% Tween 20 for 15 minutes. 8. Rinse the nitrocellulose in 4-5 changes of water with a final wash in water for 2 hours at RT with agitation. 9. Use detection method of choice (enhanced chemiluminescence was used).

Hello Abcam, This antibody is an unpurified cell culture supernant, so the antibody concentration can not be determined against the background of other proteins. The lack of a known concentration is compensated for by stated specific dilutions for suggested applications. The antibody is recommended for immunoprecipitation citing a specificic reference: Gotzmann, J., et al., Electrophoresis 18: 2645–2653, 1997. An epitope determination has not been performed.