E.coli-derived human Alpha Actinin 4 recombinant protein (Position: E561-V661). Human Alpha Actinin 4 shares 99% and 98% amino acid (aa) sequence identity with mouse and rat Alpha Actinin 4, respectively.
Application Notes: Western blot, 0.1-0.5µg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5µg/ml, Human Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human Flow Cytometry (Fixed), 1-3 µg/1x106 cells, Human, Mouse, Rat. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
Flow Cytometry analysis of C6 cells using anti-Alpha Actinin 4 antibody. Overlay histogram showing C6 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 rabbit anti-Alpha Actinin 4 Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Flow Cytometry analysis of HEPA1-6 cells using anti-Alpha
Flow Cytometry analysis of HEPA1-6 cells using anti-Alpha Actinin 4 antibody. Overlay histogram showing HEPA1-6 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 rabbit anti-Alpha Actinin 4 Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Flow Cytometry analysis of U87 cells using anti-Alpha Actinin 4 antibody. Overlay histogram showing U87 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 rabbit anti-Alpha Actinin 4 Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
IF analysis of Alpha Actinin 4 using anti-Alpha Actinin 4 antibody. Alpha Actinin 4 was detected in an immunocytochemical section of Hela 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 5 µg/mL rabbit anti-Alpha Actinin 4 Antibody overnight at 4C. DyLight488 Conjugated Goat Anti-Rabbit 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 Alpha Actinin 4 using anti-Alpha Actinin 4 antibody. Alpha Actinin 4 was detected in a paraffin-embedded section of human colonic adenocarcinoma 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 2 µg/ml rabbit anti-Alpha Actinin 4 Antibody overnight at 4C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
IHC analysis of Alpha Actinin 4 using anti-Alpha Actinin 4 antibody. Alpha Actinin 4 was detected in a paraffin-embedded section of human prostate 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 2 µg/ml rabbit anti-Alpha Actinin 4 Antibody overnight at 4C. Peroxidase Conjugated Goat Anti-rabbit IgG was used
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