Western blotting analysis using High mobility group box 1 antibody (Cat#1114). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with High mobility group box 1 antibody (Cat#1114, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Immunohistochemistry was performed on paraffin-embedded human sigmoid colon carcinoma using high mobility group box 1 antibody (Cat#1114, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar, 25 μm.
Immunohistochemistry was performed on paraffin-embedded mouse heart using high mobility group box 1 antibody (Cat#1114, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar, 25 μm.
Immunohistochemistry was performed on paraffin-embedded mouse kidney using high mobility group box 1 antibody (Cat#1114, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar, 25 μm.
Immunohistochemistry was performed on paraffin-embedded mouse brain using high mobility group box 1 antibody (Cat#1114, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar, 25 μm.
Immunohistochemistry was performed on paraffin-embedded mouse skeletal muscle using high mobility group box 1 antibody (Cat#1114, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar, 25 μm.
Immunohistochemistry was performed on paraffin-embedded mouse liver using high mobility group box 1 antibody (Cat#1114, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar, 25 μm.
Immunohistochemistry was performed on paraffin-embedded mouse lung using high mobility group box 1 antibody (Cat#1114, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar, 25 μm.
Immunocytochemical staining of HepG2 cells with High mobility group box 1 antibody (Cat#1114, 1:1,000). Nuclei were stained blue with DAPI; High mobility group box 1 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: High. Scale bar, 20 μm.
Flow cytometric analysis of High mobility group box 1 expression in HepG2 cells using High mobility group box 1 antibody (Cat#1114, 1:2,000). Green, isotype control; red, High mobility group box 1.