Anti-Desmoglein 2 antibody, MBS175420, Western blotting
All lanes: Anti Desmoglein 2 (MBS175420) at 0.5ug/ml
Lane 1: HT1080 Whole Cell Lysate at 40ug
Lane 2: HELA Whole Cell Lysate at 40ug
Lane 3: SW620 Whole Cell Lysate at 40ug
Lane 4: SCG Whole Cell Lysate at 40ug
Lane 5: COLO320 Whole Cell Lysate at 40ug
Predicted bind size: 122KD
Observed bind size: 122KD
Western blot analysis of DSG2 using anti-DSG2 antibody (MBS175420). Electrophoresis was performed on a 8% SDS-PAGEgel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours.The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human A549- WT whole cell lysates, Lane 2: human A549-DSG2 KO whole cell 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-DSG2 antigen affinity purified polyclonal antibody (MBS175420) at 0.5 ug/mL overnight at 4°C, 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 ECL Plus Western Blotting Substrate with Tanon 5200 system. A specific band was detected for DSG2 at approximately 160 kDa. The expected band size for DSG2 is at 122 kDa.
Western blot analysis of DSG2 using anti-DSG2 antibody (MBS175420). 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 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: human A549 whole cell lysates, Lane 4: rat heart tissue lysates, Lane 5: 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-DSG2 antigen affinity purified polyclonal antibody (Catalog # MBS175420) at 0.5 ug/mL overnight at 4°C, 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 DSG2 at approximately 160 kDa. The expected band size for DSG2 is at 122 kDa
IHC analysis of DSG2 using anti-DSG2 antibody (MBS175420). DSG2 was detected in paraffin-embedded section of human breast carcinoma 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 1ug/ml rabbit anti-DSG2 Antibody (MBS175420) overnight at 4°CC. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of DSG2 using anti-DSG2 antibody (MBS175420). DSG2 was detected in paraffin-embedded section of rat intestine 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 1ug/ml rabbit anti- DSG2 Antibody (MBS175420) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IF analysis of DSG2 using anti-DSG2 antibody (MBS175420). DSG2 was detected in immunocytochemical section of MCF-7 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 2ug/mL rabbit anti-DSG2 Antibody (PA1559) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Flow Cytometry analysis of 293T cells using anti-DSG2 antibody (MBS175420).Overlay histogram showing 293T cells stained with PA1559 (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-DSG2 Antibody (PA1559,1ug/1x106 cells) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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