Figure 1. Western blot analysis of M Cadherin/CDH15 using anti-M Cadherin/CDH15 antibody (MBS1755770).
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 293T whole cell lysates,
Lane 3: rat L6 whole cell lysates,
Lane 4: mouse C2C12 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-M Cadherin/CDH15 antigen affinity purified polyclonal antibody (#MBS1755770) at 0.5 ug/mL overnight at 4 degree 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 M Cadherin/CDH15 at approximately 130 kDa. The expected band size for M Cadherin/CDH15 is at 89 kDa.
Figure 2. IHC analysis of M Cadherin/CDH15 using anti-M Cadherin/CDH15 antibody (MBS1755770).
M Cadherin/CDH15 was detected in a paraffin-embedded section of mouse skeletal muscle tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-M Cadherin/CDH15 Antibody (MBS1755770) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Figure 3. IHC analysis of M Cadherin/CDH15 using anti-M Cadherin/CDH15 antibody (MBS1755770).
M Cadherin/CDH15 was detected in a paraffin-embedded section of rat skeletal muscle tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-M Cadherin/CDH15 Antibody (MBS1755770) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Figure 4. Flow Cytometry analysis of C2C12 cells using anti-M Cadherin/CDH15 antibody (MBS1755770).
Overlay histogram showing C2C12 cells stained with MBS1755770 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-M Cadherin/CDH15 Antibody (MBS1755770, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG (MBS177405, 5-10 ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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