NPTII antibody (20C-CR1112RP)
Rabbit polyclonal NPTII antibody
|Synonyms||Polyclonal NPTII antibody, Anti-NPTII antibody, Neomycin Phosphotransferase II antibody|
|Cross Reactivity||To be determined by end-user|
|Immunogen||NPTII antibody was raised in rabbit using purified neomycin phosphotransferase II as the immunogen.|
|Form & Buffer||Supplied as an IgG fraction in PBS, pH 7.4 with 0.05% NaN3|
Usage & Assay Information
|Usage Recommendations||Use at starting dilution 1:500|
Storage & Safety
|Storage||Store at 4 deg C for short term storage. Aliquot and store at -80 deg C for long term storage. Avoid repeated freeze/thaw cycles.|
|Biological Significance||Neomycin phosphotransferase II confers resistance to kanamycin and neomycin in bacteria and G418 in mammalian cells. These antibiotics bind to ribosomal components and inhibit protein synthesis. NPT II inhibits these antibiotics through phosphorylation, which is thought to interfere with their active transport into the cell. Immunoblot analysis of transfected cell lysates with anti NPT II can be used to gauge relative transfection efficiencies in experiments where multiple transfections are performed in parallel with different plasmids.|
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|Submitted By||Nico Meyer|
|Date||16th March 2015|
|Application product used in||Western Blotting|
|Usage||1:1000 in 3% milk for 1 hour|
|Comments||Excellent antibody, specific and clear, one of the few available ones that works!
Samples were isolated from plant extract from Arabidopsis seedlings. 100ug protein was loaded onto the gel and transferred to a PVDF membrane for blotting. NPTII antibody was dluted 1:1000 in 3% milk and allowed to incubate for 1 hour.
Antibody works with high specificity on multiple samples containing crude extract of transgenic Arabidopsis thaliana. We have used the following lines expressing E. coli NPTII to test antibody specificity: ku70 (Riha et al. (2002)), prt6-2 (SALK_051088, aka prt6-5 in Graciet et al. (2009), and prt6-1 in Holman et al. (2009)), and mpk1 (Bartels et al. (2009)). A plant sensitive for kanamycin was used as negative control.