Cell lysates were separated on Bolt 4–12% Bis-Tris Plus (Novex) gradient gels. Proteins were transferred onto 0.45 μm nitrocellulose membranes (Amersham Protran) using Trans-blot SD cell (Bio-Rad). The membrane was blocked using PBS containing 0.1% Tween 20 and 5% Marvel milk powder for 1 h at room temperature. The membrane was incubated with either anti-RVFV N polyclonal rabbit antibody (1:5000) [23 (link)] or beta-actin rabbit monoclonal antibody (1:1000; Cell Signalling Technology) overnight at 4°C. A horseradish peroxidase (HRP) anti-rabbit secondary antibody (1:1000) was then applied to the membrane. Pierce ECL Western Blotting Substrate (Thermo-Scientific) followed by visualisation on a Biorad ChemiDoc system.
Beta actin rabbit monoclonal antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Beta-actin rabbit monoclonal antibody is a laboratory tool used to detect and quantify the presence of the beta-actin protein in biological samples. It is a highly specific and sensitive reagent that binds to the beta-actin protein, allowing researchers to study its expression and localization in cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Trans blot sd cell, by Bio-Rad (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Chemidoc system, by Bio-Rad (1 mentions)
Goat anti rabbit igg hrp sc 2030, by Santa Cruz Biotechnology (1 mentions)
Pierce ecl western blotting substrate, by Bio-Rad (1 mentions)
Chemidoc mp imaging system, by Bio-Rad (1 mentions)
Hrp conjugated goat anti rabbit antibody, by Cell Signaling Technology (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Mouse monoclonal opn antibody, by Santa Cruz Biotechnology (1 mentions)
4 protocols using beta actin rabbit monoclonal antibody
1
Western Blot Analysis of RVFV N Protein
2
Western Blot of Histone Modifications
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Cell lysates were collected from GM12878 and LCL8664 cells in RIPA buffer supplemented with protease inhibitors. Protein content was quantified using the Pierce BCA Protein Assay Kit, and samples were prepared for gel loading (25 μg protein, 5% BME, 6x SDS buffer) and boiled at 95°C for 5 min. Samples were run on a 4–20% Mini-PROTEIN TGX Precast Protein Gel at 100V for 90 min, transferred to a PVDF for 2 h at 25V using the XCell II semi-wet transfer system and 20% methanol transfer buffer. The membrane was blocked with 5% milk in TBS-T overnight at 4°C, incubated with primary antibody (1:1000 dilution in 5% milk) for 2 h at room temperature, washed 3x with TBS-T, and incubated with secondary antibody for 1 h at room temperature. The membrane was washed 3x with TBS-T prior to incubation with Pierce ECL Western Blotting Substrates and imaged using the colorimetric and chemiluminescence channels of the BioRad ChemiDoc MP imaging system. Primary antibody (Rabbit polyclonal H3K27ac antibody, Abcam ab4729; Histone H3 rabbit monoclonal antibody, Cell Signaling #4499; IRF4 rabbit polyclonal antibody, Cell Signaling #4964; beta-actin rabbit monoclonal antibody, Cell Signaling #8457). Secondary antibody (Goat anti-rabbit IgG-HRP, sc-2030, Santa Cruz Biotechnology).
3
Seladin-1 Expression in RT4 Cells
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RT4 cells were seeded at a concentration of 2.5 × 105 cells/ml on a tissue culture dish and incubated for 24 hrs. Next, the cells were treated with goniothalamin at the concentration of 61 μM (IC50) for 2, 4, 8, 16, and 24 hrs. Following treatment, cell lysates were collected and protein samples were prepared and denatured at 95°C for 5 mins. Next, 20 μg/ml lysate was resolved on 12% SDS-polyacrylamide gel electrophoresis and blotted onto a polyvinylidene fluoride membrane. Western blotting was carried out as described previously [18 (link)]. Rabbit monoclonal anti-Seladin-1 antibody (Sigma, USA) was used at a dilution of 1:5000 in 5% skimmed milk and incubated at 4°C overnight. Meanwhile, rabbit monoclonal beta actin antibody (Cell Signaling, USA) was used at a dilution of 1:10000. After incubation of the primary antibody, secondary antibody (goat anti-rabbit HRP conjugated antibody, Cell Signaling, USA) was added and incubated for 1 hr. prior to detection by chemiluminenscene (Amersham, UK).
4
Protein Expression Analysis in Tissues and Cells
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Tissues and cultured cells proteins were extracted using lysis buffer supplemented and protein concentrations were measured using BCA Protein Assay Kit. Protein was loaded at the concentration of 1000 ng/per well on SDS-polyacrylamide gels and transferred to polyvinylidene fluoride membranes (Millipore) Membranes were blocked with 5% skim milk for 1 h at room temperature. Membranes were incubated with primary antibodies in 5% BSA (1:1000) as follows: mouse monoclonal OPN antibody (Santa Cruz Biotechnology, sc-21742), mouse monoclonal insulin-like growth factor-binding protein-1 (IGFBP-1) antibody (Santa Cruz, sc-25257), mouse monoclonal vascular endothelial growth factor A (VEGFA) antibody (Santa Cruz, sc-365578) and rabbit monoclonal beta-ACTIN antibody (Cell Signaling Technology, 4970) overnight and washed. Then membranes were incubated with horseradish peroxidaselabeled anti-mouse or anti-rabbit IgG secondary antibody in 5% skim milk (1:4000) at room temperature for 1 h and visualized by enhanced chemiluminescence.
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