Imagej

Manufactured by Vilber

ImageJ is an open-source image processing and analysis software. It provides a range of tools for viewing, editing, analyzing, and processing digital images.

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Lab products found in correlation

Bio1d, by Vilber (2 mentions) Fusion solo, by Vilber (2 mentions) Protease inhibitor cocktail, by Roche (1 mentions)

2 protocols using imagej

Immunoprecipitation and Western Blot Workflow

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For immunoprecipitation and Western blot, cells were lysed on ice for 30 min with lysis buffer (500 mM Tris–HCl pH 7.4, 2 mM benzamidine, 10 mM NaF, 20 mM EDTA, 0.5% NP40 and a protease inhibitor cocktail (Roche, CH)). The lysate was then clarified by centrifugation at 4 °C for 3 min at 5000 rpm. Lysates were pre-cleared using Sepharose G-beads only for 30 min at 4 °C before immunoprecipitation (G-beads plus antibody) turning on a wheel overnight at 4 °C. The beads were then washed 3× with lysis buffer before adding 4× Sample Buffer including beta-mercaptoethanol. The samples were boiled 5 min at 95 °C and vortexed before loading and migrating on 4–12% or 4–20% Tris-glycine SDS-PAGE gels. Blots were revealed using a Fusion Solo (Vilber Lourmat, CH) and quantified with ImageJ or Bio1D (Vilber Lourmat, CH).

Sandoz P.A., Denhardt-Eriksson R.A., Abrami L., Abriata L.A., Spreemann G., Maclachlan C., Ho S., Kunz B., Hess K., Knott G., S. Mesquita F., Hatzimanikatis V, & van der Goot F.G. (2023). Dynamics of CLIMP-63 S-acylation control ER morphology. Nature Communications, 14, 264.

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Immunoprecipitation and Western Blot Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

For immunoprecipitation and Western blot, cells were lysed on ice for 30min with lysis buffer (500 mM Tris-HCl pH 7.4, 2 mM benzamidine, 10 mM NaF, 20 mM EDTA, 0.5% NP40 and a protease inhibitor cocktail (Roche, CH)). The lysate was then clari ed by centrifugation at 4°C for 3 min at 5000 rpm. Lysates were pre-cleared using Sepharose G-beads only for 30 min at 4°C before immunoprecipitation (Gbeads plus antibody) turning on a wheel overnight at 4°C. The beads were then washed 3x with lysis buffer before adding 4x Sample Buffer including beta-mercaptoethanol. The samples were boiled 5 min at 95°C and vortexed before loading and migrating on 4-12% or 4-20% Tris-glycine SDS-PAGE gels.
Blots were revealed using a Fusion Solo (Vilber Lourmat, CH) and quanti ed with ImageJ or Bio1D (Vilber Lourmat, CH).

Sandoz P.A., Denhardt-Eriksson R., Abrami L., Abriata L.A., Spreemann G., Maclachlan C., S H., Kunz B., Hess K., Knott G., Mesquita F.S., Hatzimanikatis V., & Goot F.G.v.d. (2022). Model-driven understanding of CLIMP-63 structure and S-acylation reveals fine tuning of ER morphology.

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