Western blotting analysis using creatine kinase B type antibody (Cat#63170). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with creatine kinase B type antibody (Cat#63170, 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).
Western blotting analysis using creatine kinase B type antibody (Cat#63170). Creatine kinase B type expression in wild-type (WT) and creatine kinase B type(CKB) shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with creatine kinase B type antibody (Cat#63170, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Immunohistochemistry was performed on paraffin-embedded human sigmoid colon carcinoma using creatine kinase B type antibody (Cat#63170, 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 Hela cells with creatine kinase B type antibody (Cat#63170, 1:1,000). Nuclei were stained blue with DAPI; Creatine kinase B type 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.
Immunocytochemical staining of HeLa cells using creatine kinase B type antibody (Cat#63170, 1:1,000), Top panel: wild-type (WT); Bottom panal: Creatine kinase B type shRNA knockdown (KD). Nuclei were stained blue with DAPI; Creatine kinase B was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm. Permeabilization: Triton.
Flow cytometric analysis of creatine kinase B type expression in HeLa cells using creatine kinase B type antibody (Cat#63170, 1:2,000). Green, isotype control; red, Creatine kinase B type.
Validation of creatine kinase B type knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with creatine kinase B type antibody (Cat#63170, 1:2,000) and analyzed using BD flow cytometer.
