Western blot, 0.1-0.25µg/ml, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1µg/ml, Mouse, Rat Flow Cytometry (Fixed), 1-3µg/1x10 6 cells, Mouse ELISA, 0.1-0.5µg/ml, - Rat
Flow Cytometry analysis of mouse spleen tissues using anti-CD3 epsilon/Cd3e antibody. Overlay histogram showing mouse spleen tissues (Blue line). The tissues were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-CD3 epsilon/Cd3e 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 CD3 epsilon/Cd3e using anti-CD3 epsilon/Cd3e antibody. CD3 epsilon/Cd3e was detected in a paraffin-embedded section of mouse lymph node 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-CD3 epsilon/Cd3e 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 CD3 epsilon/Cd3e using anti-CD3 epsilon/Cd3e antibody. CD3 epsilon/Cd3e was detected in a paraffin-embedded section of mouse spleen 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-CD3 epsilon/Cd3e 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 CD3 epsilon/Cd3e using anti-CD3
IHC analysis of CD3 epsilon/Cd3e using anti-CD3 epsilon/Cd3e antibody. CD3 epsilon/Cd3e was detected in a paraffin-embedded section of rat lymph node 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-CD3 epsilon/Cd3e 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.
Western blot analysis of CD3 epsilon/Cd3e using anti-CD3 epsilon/Cd3e 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 30 ug of sample under reducing conditions. Lane 1: rat thymus tissue lysates, Lane 2: mouse thymus 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-CD3 epsilon/Cd3e antigen affinity purified polyclonal antibody at 0.25 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 CD3 epsilon/Cd3e at approximately 23 kDa. The expected band size for CD3 epsilon/Cd3e is at 21 kDa.
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