Figure 1. Western blot analysis of SEPT6/SEPTIN6 using anti-SEPT6/SEPTIN6 antibody (MBS1755772).
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 SH-SY5Y whole cell lysates,
Lane 2: human MOLT-4 whole cell lysates,
Lane 3: rat thymus tissue lysates,
Lane 4: rat brain tissue lysates,
Lane 5: mouse brain 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-SEPT6/SEPTIN6 antigen affinity purified polyclonal antibody (#MBS1755772) at 0.25 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 SEPT6/SEPTIN6 at approximately 55 kDa. The expected band size for SEPT6/SEPTIN6 is at 50 kDa.
Figure 2. IHC analysis of SEPT6/SEPTIN6 using anti-SEPT6/SEPTIN6 antibody (MBS1755772).
SEPT6/SEPTIN6 was detected in a paraffin-embedded section of human spleen 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-SEPT6/SEPTIN6 Antibody (MBS1755772) 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 SEPT6/SEPTIN6 using anti-SEPT6/SEPTIN6 antibody (MBS1755772).
SEPT6/SEPTIN6 was detected in a paraffin-embedded section of human tonsil 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-SEPT6/SEPTIN6 Antibody (MBS1755772) 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 ANA-1 cells using anti-SEPT6/SEPTIN6 antibody (MBS1755772).
Overlay histogram showing ANA-1 cells stained with MBS1755772 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SEPT6/SEPTIN6 Antibody (MBS1755772, 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.
Figure 5. Flow Cytometry analysis of HEL cells using anti-SEPT6/SEPTIN6 antibody (MBS1755772).
Overlay histogram showing HEL cells stained with MBS1755772 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SEPT6/SEPTIN6 Antibody (MBS1755772, 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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