Protein levels were determined using Western blot analysis following the protocol in the lab (Ni et al., 2012 (link)) with modification. The primary antibodies (Cell Signaling Technology) against STAT3 (1:2000), p-STAT3 (1:2000), JAK-2 (1:2000), p-JAK-2 (1:2000), Cleaved caspase-3(1:2000), Cleaved caspase-9 (1:2000), Bax (1:2000), Bcl-2(1:2000), LC3(1:2000) and β-actin (1:2000) were used, and the HRP-conjugated anti-rabbit/mouse IgG secondary antibody (1:2000, Cell Signaling Technology) was used. Intensities of the bands were measured by Image Lab software (Bio-Rad) to assess relative protein levels.
Hrp conjugated anti rabbit mouse igg secondary antibody
Manufactured by Cell Signaling Technology
The HRP-conjugated anti-rabbit/mouse IgG secondary antibody is a laboratory reagent used in various immunoassay techniques, such as Western blotting and enzyme-linked immunosorbent assay (ELISA). This product is designed to detect and visualize primary antibodies that are raised against rabbit or mouse immunoglobulin G (IgG) proteins. The secondary antibody is conjugated with horseradish peroxidase (HRP), an enzyme that catalyzes a colorimetric or chemiluminescent reaction, enabling the detection and quantification of the target protein.
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2 protocols using hrp conjugated anti rabbit mouse igg secondary antibody
1
Western Blot Analysis of JAK-STAT Pathway
2
Protein Quantification in Lung Tissues
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Protein was isolated from lung tissues after 3 d model establishment by using the protein purification kit (Sangon, Shanghai, China). Protein levels were determined using Western blot analysis following the protocol in the lab with modification [20 , 21 (link)]. The primary antibodies (Cell Signaling Technology) against p-STAT3 (1 : 2000), p-JAK-2 (1 : 2000), STAT3 (1 : 2000), JAK-2 (1 : 2000), caspase 3 (1 : 2000), BAX (1 : 2000), Bcl-2 (1 : 2000), and β-actin (1 : 2000) were used, and the HRP-conjugated anti-rabbit/mouse IgG secondary antibody (1 : 2000, Cell Signaling Technology) was used. Intensities of the bands were measured by Image Lab software (Bio-Rad) to assess relative protein levels.
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