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What were the Western blot conditions used for the blot on the datasheet? |
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ANSWER: |
Thank you for your enquiry. The membrane used was 0.45micron nitrocellulose, blocked in 5% non fat milk in TBS, antibody incubations in TBS +0.1% tween 20, ab7255 conc was equivalent of 1:500 from our standard tissue culture supernatant prep and incubation time was 1hr at RT with shaking. Detection in this case was with Goat anti-mouse alkaline phosphatase conjugate and development was with BCIP/NBT. The blot was made quite a while ago, probably about 1993, and if we were doing it today we would use chemiluminesnce and probably 1:1,000 to 1:2,000 ab concentration. I hope this information helps. Please do not hesitate to contact us again if you need anything further. |
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Hi, I'm interested in finding a good NF-L antibody. I am curious about the amount you reccomend using for immunoblotting (i.e. what kind of dilution, 1:1000, 1:200, 1:5000, etc. etc.) Normally the data sheet indicates this information, but I did not see it there. Thanks. |
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ANSWER: |
Dear Sir/Madam: Our anti-NF-L monoclonal antibody (catalog# NB 300-132) isunpurified mouse ascites (50 ul). We suugest a working dilution of 1:1,000 for ECL. Best Regards, Kate Lynott |
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 68kDa Neurofilament antibody [DA2] - Neuronal Marker (ab7255)](/ps/datasheet/images/7/ab7255/68kDa-Neurofilament-Primary-antibodies-ab7255-1.jpg)