Western blot, 0.25-0.5µg/ml, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1µg/ml, Human Immunocytochemistry/Immunofluorescence, 2µg/ml, Human Flow Cytometry (Fixed), 1-3µg/1x10 6 cells, Human
Flow Cytometry analysis of A431 cells using anti-MORC3 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 rabbit anti-MORC3 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.
IHC analysis of MORC3 using anti-MORC3 antibody.
IF analysis of MORC3 using anti-MORC3 antibody. MORC3 was detected in immunocytochemical section of A431 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 rabbit anti-MORC3 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 MORC3 using anti-MORC3 antibody. MORC3 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 rabbit anti-MORC3 Antibody overnight at 4C. Biotinylated goat anti-rabbit 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 MORC3 using anti-MORC3 antibody. MORC3 was detected in paraffin-embedded section of human rectal 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 rabbit anti-MORC3 Antibody overnight at 4C. Biotinylated goat anti-rabbit 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 MORC3 using anti-MORC3 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: rat heart tissue lysates, Lane 2: rat liver tissue lysates, Lane 3: mouse heart tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MORC3 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for MORC3 at approximately 120 KD. The expected band size for MORC3 is at 107 KD.
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