Proteins were extracted with radioimmunoprecipitation assay (RIPA) lysis buffer and boiled for denaturation. Sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) was performed on samples containing 30 µg of protein using a 10% polyacrylamide gel. After electrotransfer and blocking with 3% bovine serum albumin, the membranes were incubated overnight at 4 °C with primary antibodies. Rabbit antibodies against TFEB, HLA-A, GAPDH, Histone-H3 (1:2000; Proteintech, China), PD-L1, PD-L2 (1:5000; Abcam, USA), and mouse antibody against β-actin (1:5000; Proteintech) were used as primary antibodies. After incubating with anti-rabbit or anti-mouse secondary antibody (1:5000; Proteintech) for 60 min at room temperature, enhanced chemiluminescence (ECL, NJ, USA) was used to visualize the bands.
Mouse antibody against β actin
Manufactured by Proteintech
Sourced in United States
The mouse antibody against β-actin is a laboratory reagent used for the detection and quantification of the β-actin protein, a common housekeeping gene expressed in most cell types. This antibody can be used in various immunoassay techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to identify and measure the levels of β-actin in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti rabbit or anti mouse secondary antibody, by Proteintech (1 mentions)
Pd l2, by Abcam (1 mentions)
Pd l1, by Abcam (1 mentions)
Gapdh, by Proteintech (1 mentions)
Histone h3, by Proteintech (1 mentions)
Anti mouse hrp conjugated secondary antibody, by Bio-Rad (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
2 protocols using mouse antibody against β actin
1
Western Blot Analysis of Immune Markers
2
Mutation Analysis using Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Primers carrying the mutations were designed using NEBaseChanger software.
Primer sequences: I80P-F: 5'-TTTCCTTATCCCATTTCTGGTGCAG-3', I80P-R: non-fat dry milk in Tris-buffer saline with 0.1% Triton X-100 (TBST) for 2 hours at room temperature. Then, the membranes were incubated with the primary antibodies in blocking solution overnight at 4°C. The following primary antibodies were used for the Western blotting: mouse antibodies against Flag (Sigma-Aldrich, St. Louis, MO, USA)
and mouse antibody against β-actin (Proteintech Group, Chicago, IL, USA). The primary antibodies were detected with anti-mouse HRP-conjugated secondary antibodies (1:5000; Bio-Rad, Hercules, CA, USA), and the signal was developed using Supersignal West Pico Chemiluminescent Substrate (Thermo Scientific). The relative intensity of the immunoreactive bands was quantified using the gel analysis tool provided in the ImageJ software. Normalization of the proteins of interest was performed relative to β-actin. Mean fluorescence intensity was calculated for each group.
Primer sequences: I80P-F: 5'-TTTCCTTATCCCATTTCTGGTGCAG-3', I80P-R: non-fat dry milk in Tris-buffer saline with 0.1% Triton X-100 (TBST) for 2 hours at room temperature. Then, the membranes were incubated with the primary antibodies in blocking solution overnight at 4°C. The following primary antibodies were used for the Western blotting: mouse antibodies against Flag (Sigma-Aldrich, St. Louis, MO, USA)
and mouse antibody against β-actin (Proteintech Group, Chicago, IL, USA). The primary antibodies were detected with anti-mouse HRP-conjugated secondary antibodies (1:5000; Bio-Rad, Hercules, CA, USA), and the signal was developed using Supersignal West Pico Chemiluminescent Substrate (Thermo Scientific). The relative intensity of the immunoreactive bands was quantified using the gel analysis tool provided in the ImageJ software. Normalization of the proteins of interest was performed relative to β-actin. Mean fluorescence intensity was calculated for each group.
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