Western blot analysis was performed at 24 hours after MCAo from the ipsilateral cortex and subcortical regions. Brain protein abundance was determined by Bio‐Rad Assay Method. A total of 20‐25 μg of equivalent protein was loaded on NuPAGE 4%‐12% Bis‐Tris gels and run at 125 V for 120 minutes, and then, proteins were transferred onto PVDF (Invitrogen) paper. TREM2 and MANF proteins were detected using anti‐TREM2 (Millipore‐Sigma) and anti‐MANF antibody (Abcam). Finally, TREM2 and MANF proteins were imaged in Fujifilm LAS‐3000 using Amersham™ ECL™ Western Blotting Detection Reagents. TREM 2 and MNAF protein bands were quantified by Multigage software, and the results were expressed as TREM2/GAPDH and MANF/GAPDH ratio, respectively.
Las 3000
Manufactured by Cytiva
The LAS-3000 is a versatile laboratory imaging system designed for a range of applications. It features a high-resolution camera, adjustable lighting, and software for image capture and analysis. The LAS-3000 is a tool for researchers and scientists to document and analyze their experimental results.
Automatically generated - may contain errors
Lab products found in correlation
Supersignal west pico plus chemiluminescent substrate, by Thermo Fisher Scientific (2 mentions)
Horseradish peroxidase conjugated goat anti rabbit secondary antibody, by Thermo Fisher Scientific (2 mentions)
Amersham imager 600, by Cytiva (2 mentions)
Polyvinylidene fluoride (pvdf), by Thermo Fisher Scientific (1 mentions)
Anti manf antibody, by Abcam (1 mentions)
Amersham ecl western blotting detection reagent, by Cytiva (1 mentions)
3 protocols using las 3000
1
Quantifying TREM2 and MANF Proteins in Stroke
2
Immunoblotting Protein Analysis
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SDS-PAGE analysis of proteins was performed as described [67] . Tween-20 and incubated with 1/5,000 diluted goat anti-rabbit horseradish peroxidase-conjugated secondary antibody (Thermo Scientific) in 5% milk TBS for 1 hour at room temperature with shaking. The membrane was then washed 4 times with 1X TBS with 0.1% (v/v) Tween-20. The antibody-antigen complex was detected by using the SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Scientific) and the FujiFilm LAS-3000 or Amersham imager 600 analyzers.
3
Immunoblotting Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SDS-PAGE analysis of proteins was performed as described [67] . Tween-20 and incubated with 1/5,000 diluted goat anti-rabbit horseradish peroxidase-conjugated secondary antibody (Thermo Scientific) in 5% milk TBS for 1 hour at room temperature with shaking. The membrane was then washed 4 times with 1X TBS with 0.1% (v/v) Tween-20. The antibody-antigen complex was detected by using the SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Scientific) and the FujiFilm LAS-3000 or Amersham imager 600 analyzers.
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