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Pneumococcal serotypes were identified using polyclonal rabbit antisera (Statens Seruminstitut, Copenhagen, Denmark).
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Enteroendocrine cells were identified using rabbit polyclonal antibodies to Chromogranin A (Abcam, Cambridge, UK), and we used rabbit polyclonal antibodies to CD117 (Dako, Glostrup, Denmark) for mast cells, rabbit polyclonal antibodies to CD68 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for macrophages and rabbit monoclonal antibodies to CD11c (Abcam, Cambridge, UK) for dendritic cells.
CRF (25 kDa) was identified using primary rabbit polyclonal antibody (CRF, FL-196 antibody, Santa Cruz Biotechnology, dilution 1 : 500).
HIF-1 α (120 kDa) was identified using primary rabbit polyclonal antibody (HIF-1 α, H-206X antibody, Santa Cruz Biotechnology, dilution 1 : 300).
Actin was identified using a rabbit polyclonal IgG (anti-actin N-terminal antibody produced in rabbit 1 750, 12 hours, 4°C) (Sigma-Aldrich) and a secondary (H+L -HRP conjugated goat anti-rabbit IgG (1:2,000, 80 minutes, 22°C) (Bio-Rad Laboratories, Inc).. CH+L -HRPnesconjugatedction was performed as described above, exposingoate film to the membrane for 2 minutes.
CTF- γ cleavage products (as well as CTF-α and CTF-beta) were resolved electrophoretically in 10 20% Tris-Tricine gels (Bio-Rad, CA), and identified using the anti APP polyclonal C8 antibody.
Following 7 days of culture, matrices were re-stained selectively for fibronectin using a polyclonal antibody and nuclei were identified using 4',6-diamidino-2-phenylindole (DAPI).
Proteins were identified using anti-VCC [19] and anti-PrtV [10] polyclonal rabbit antisera at a final dilution of 1∶10,000 and 1∶200,000 respectively.
Lysosome or ER-enriched membrane fractions were identified, using a monoclonal mouse anti-LAMP-1 (Abcam) and a polyclonal rabbit anti-calnexin antibody (Stressgen, Ann Arbor, MI, USA).
ED-A FN was identified using the 3E2 monoclonal IgM antibody (Sigma) [ 30] and a rabbit polyclonal antibody recognizing lysyl oxidase (LOX) was used to identify cells synthesizing collagen and elastin [ 31].
Lipid droplets were identified using either a 12 µg/mL Nile Red solution or by using the lipid droplet specific rabbit polyclonal LSD2-3b at 1∶500 and a goat-anti-rabbit Alexa 568 at 1∶1000 as the secondary.
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