E. coli-derived human Calpain 2 recombinant protein (Position: R500-L700). Human Calpain 2 shares 93.5% and 92.5% amino acid (aa) sequence identity with mouse and rat Calpain 2, respectively.
Application Notes: Western blot, 0.1-0.5µg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1µg/ml Immunocytochemistry/Immunofluorescence, 2µg/ml Flow Cytometry (Fixed), 1-3µg/1x106 cells. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
Flow Cytometry analysis of A431 cells using anti-Calpain 2 antibody. Overlay histogram showing A431 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Calpain 2 Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-mouse IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Flow Cytometry analysis of U87 cells using anti-Calpain 2 antibo
IF analysis of Calpain 2 using anti-Calpain 2 antibody. Calpain 2 was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL mouse anti-Calpain 2 Antibody overnight at 4C. DyLight488 Conjugated Goat Anti-mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
IF analysis of Calpain 2 using anti-Calpain 2 antibody. Calpain 2 was detected in immunocytochemical section of U251 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL mouse anti-Calpain 2 Antibody overnight at 4C. DyLight488 Conjugated Goat Anti-mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
IHC analysis of Calpain 2 using anti-Calpain 2 antibody. Calpain 2 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-Calpain 2 Antibody overnight at 4C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of Calpain 2 using anti-Calpain 2 antibody. Calpain 2 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-Calpain 2 Antibody overnight at 4C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of Calpain 2 using anti-Calpain 2 antibody. Calpain 2 was detected in paraffin-embedded section of mouse lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-Calpain 2 Antibody overnight at 4C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Western blot analysis of Calpain 2 using anti-Calpain 2 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: A549 whole cell lysates, Lane 2: U20S whole cell lysates, Lane 3: COS-7 whole
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