Hippocampal tissue was prepared for protein extraction with RIPA buffer (Bioteke, China). Protein concentrations in the homogenates were determined with the micro BCA protein assay kit (Bioteke, China) with an enzyme-labelling instrument (Thermo varioskan flash, USA). Approximately 40 μg of total protein samples were loaded per lane onto a 10% SDS-polyacrylamide gel (Beyotime, China). Proteins were transferred to 0.45 μm polyvinylidene fluoride membranes (Millipore, USA) after electrophoretic separation. All membranes were then blocked with 5% skim milk in TBST at 37 °C for 1 h and probed with anti- NSE (at dilution of 1/2000) (Abcam, UK), anti-HDAC2 (at dilution of 1/1500) (Millipore, USA), and anti-β-actin (at dilution of 1/800) (Santa Cruz, USA) primary antibodies at 4°C overnight. The proteins were probed with HRP-conjugated secondary antibodies (at dilution of 1/5000) (Santa Cruz, USA) at room temperature for 1 h and visualised using an enhanced chemiluminescence kit (Millipore, USA) with an ECL Imaging System (SynGene GBOX, UK). Band intensities were quantified using ImageJ software.
Micro bca protein assay kit
Manufactured by BioTeke
Sourced in China
The Micro BCA Protein Assay Kit is a colorimetric detection method used to determine the concentration of protein in a sample. It utilizes bicinchoninic acid (BCA) to quantify the total protein present in the sample, with the resulting color change being measured spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Hrp conjugated secondary antibody, by Santa Cruz Biotechnology (2 mentions)
Sds polyacrylamide gel, by Beyotime (2 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Enhanced chemiluminescence kit, by Merck Group (1 mentions)
Ripa buffer, by BioTeke (1 mentions)
Anti hdac2, by Merck Group (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Anti nse, by Abcam (1 mentions)
Hatf00010, by Merck Group (1 mentions)
Ab53161, by Abcam (1 mentions)
Ab22048, by Abcam (1 mentions)
2 protocols using micro bca protein assay kit
1
Hippocampal Protein Extraction and Analysis
2
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins were extracted from the animal colonic epithelial cell monolayer and Caco-2 cells using a specific extraction kit (PP1901, BioTeke), and the protein concentrations in the homogenates were determined with a Micro BCA Protein Assay Kit (P1001, BioTeke) using an enzyme-labelling instrument (Thermo Scientific Varioskan Flash). Approximately 30 μg total protein per lane was loaded onto a 10% SDS-polyacrylamide gel (Beyotime, Haimen, China) for electrophoretic separation, after which the proteins were transferred to polyvinylidene fluoride membranes (HATF00010, Merk Millipore), which were blocked with 5% bovine serum albumin (BSA) in TBS containing 0.5% Tween for 1 h at 37 °C and then probed with anti-RARβ (ab53161, Abcam), anti-TLR4 (ab22048, Abcam), anti-ZO-2 (2847, Cell Signaling Technology) or anti-β-actin (sc-47778, Santa) primary antibodies over night at 4 °C. The proteins were subsequently probed with HRP-conjugated secondary antibodies (Santa Cruz Biotechnology) for 1 hour at room temperature and visualized on an ECL Imaging System (SynGene GBOX) using a chemiluminescence kit (1229601, Merk Millipore). Three independent experiments were performed.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!