Rabbit 800

Manufactured by LI COR

The Rabbit-800 is a laboratory instrument designed for quantitative immunoblotting and multiplex analysis. It utilizes near-infrared fluorescence technology to detect and quantify proteins separated by gel electrophoresis and transferred to a membrane. The Rabbit-800 provides high sensitivity and dynamic range for accurate protein quantification.

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Lab products found in correlation

Mouse 700, by Thermo Fisher Scientific (2 mentions) Gradient gel, by Bio-Rad (1 mentions) Rabbit tuj1, by Merck Group (1 mentions) Rabbit psd95, by Abcam (1 mentions) Odyssey detection system, by LI COR (1 mentions) Protease inhibitor cocktail tablet, by Merck Group (1 mentions) Ab15246, by Abcam (1 mentions) Odyssey imaging system, by LI COR (1 mentions) T7451, by Merck Group (1 mentions) Pvdf membrane, by Merck Group (1 mentions)

2 protocols using rabbit 800

Western Blot Analysis of Synaptic Proteins

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Slices were scraped off the membrane, treated with 2x Laemelli buffer + 10 % 2-mercaptethanol (250 μL per 3 slices) vortexed, boiled, then frozen at -20 °C until use. For use, most samples were further diluted 1:2 and loaded 8 μl per lane in a precast 4-20 % gradient gel (Bio-rad). To detect the weaker signal for PSD95, 15 μl of undiluted sample was loaded. After incubation in primary antibody overnight, blots were probed with 1:5000 mouse-700 (Life Technologies) and rabbit-800 (LI-COR) secondary antibodies then imaged using a LI-COR Odyssey detection system. Band intensity (IKK) was quantified using Odyssey software then normalised to Tuj1 signal. Primary antibodies used: mouse synaptophysin (Abcam: 1:1000), rabbit tuj1 (Sigma: 1:2500), rabbit PSD95 (Abcam: 1:500), mouse VAMP2 (Synaptic Systems: 1:10,000) and mouse RT97 (Kind gift from Dr Diane Hanger 1:500). Tuj1-normalised protein expression was then compared between WT and TgCRND8 cultures, TgCRND8 values expressed as a percentage of the WT average.

Harwell C.S, & Coleman M.P. (2016). Synaptophysin depletion and intraneuronal Aβ in organotypic hippocampal slice cultures from huAPP transgenic mice. Molecular Neurodegeneration, 11, 44.

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Quantifying Neuronal Cytoskeleton Proteins

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Following treatment, ganglia and neurites were collected and washed in PBS with complete, ethylenediaminetetraacetic acid (EDTA)-free protease inhibitor cocktail tablets (Sigma-Aldrich), and lysed directly into 2x Laemmli sample buffer. A total of 10 μL of each sample were separated on a 12% SDS-PAGE and transferred to PVDF membrane (Millipore) using the Bio-Rad Mini-PROTEAN III wet transfer system. Blots were blocked and incubated with primary antibodies overnight (in 1xTBS pH. 8.3, with 0.05% Tween 20 and 5% milk powder or 5% BSA). The antibodies were directed against α-tubulin (1/5,000; ab 15246, Abcam) and acetylated α-tubulin (1/5,000; T7451, Sigma) and detected with mouse-700 (Life Technologies) and rabbit-800 (LI-COR) secondary antibodies. Blots were then scanned and quantified using the Odyssey imaging system (LI-COR Biosciences, Lincoln, North Carolina).

Adalbert R., Kaieda A., Antoniou C., Loreto A., Yang X., Gilley J., Hoshino T., Uga K., Makhija M.T, & Coleman M.P. (2020). Novel HDAC6 inhibitors increase tubulin acetylation and rescue axonal transport of mitochondria in a model of Charcot-Marie-Tooth Type 2F. ACS chemical neuroscience, 11(3), 258-267.

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