Goat anti-Mouse IgG (H&L), HRP conjugated
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|Recommended dilution||This conjugate is suitable for all immunoassay applications. The optimal working dilution should be determined by the investigator. Suggested starting dilution(s): 1:500-1:5 000 for Immunohistochemistry 1:200-1:5000 for ELISA/Western blot|
|Selected references||Dmitrović et al. (2015). Essential oils of two Nepeta species inhibit growth and induce oxidative stress in ragweed (Ambrosia artemisiifolia L.) shoots in vitro. Acta Physiologiae Plantarum, February 2015, 37:64.|
500 femtomoles of His-tagged proteins IsiA, NifH, PsbA, PsbB and PetC were loaded per gel well in Agrisera PEB extraction buffer. Proteins were separated on 4-12 % NuPAGE PAGE Bis-Tris polycacrylamide gel (Invitrogen) and blotted 1h to PVDF. Blots were blocked with for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (goat, anti-mouse IgG horse radish peroxidase conjugated, from Agrisera AS11 1772) diluted to 1:25 000 in 2 % ECL Advance blocking reagent for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturers instructions. Exposure time was 5 seconds.
Apparent molecular weight of recombinant proteins: IsiA - 27 kda, NifH - 34 kDa, PsbA - 30-37 kDa, PsbB - 40 kDa, PetC - 23 kDa.
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