14-3-3 zeta/delta Antibody – Rabbit Polyclonal

Figure 1. Western blot analysis of 14-3-3 zeta/delta using anti-14-3-3 zeta/delta antibody (MBS1750580).
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 50ug of sample under reducing conditions.Lane 1: human Hela whole cell lysates,Lane 2: human placenta tissue lysates,Lane 3: human HepG2 whole cell lysates,Lane 4: human A549 whole cell lysates,Lane 5: human PANC-1 whole cell lysates,Lane 6: human SK-OV-3 whole cell lysates,Lane 7: human 22RV1 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-14-3-3 zeta/delta antigen affinity purified polyclonal antibody at 0.5ug/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 1:10000 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 14-3-3 zeta/delta at approximately 28KD. The expected band size for 14-3-3 zeta/delta is at 28KD.

Figure 2. Western blot analysis of 14-3-3 zeta/delta using anti-14-3-3 zeta/delta antibody (MBS1750580).
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 50ug of sample under reducing conditions.Lane 1: rat brain tissue lysates,Lane 2: rat spleen tissue lysates,Lane 3: rat lung tissue lysates,Lane 4: rat liver tissue lysates,Lane 5: mouse brain tissue lysates,Lane 6: mouse spleen tissue lysates,Lane 7: mouse lung tissue lysates,Lane 8: mouse liver tissue lysates,Lane 9: mouse kidney tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-14-3-3 zeta/delta antigen affinity purified polyclonal antibody at 0.5ug/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 1:10000 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 14-3-3 zeta/delta at approximately 28KD. The expected band size for 14-3-3 zeta/delta is at 28KD.

Figure 3. IHC analysis of 14-3-3 zeta/delta using anti-14-3-3 zeta/delta antibody (MBS1750580).
14-3-3 zeta/delta was detected in paraffin-embedded section of rat small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-14-3-3 zeta/delta Antibody (MBS1750580) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Figure 4. IHC analysis of 14-3-3 zeta/delta using anti-14-3-3 zeta/delta antibody (MBS1750580).
14-3-3 zeta/delta was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-14-3-3 zeta/delta Antibody (MBS1750580) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Figure 5. IHC analysis of 14-3-3 zeta/delta using anti-14-3-3 zeta/delta antibody (MBS1750580).
14-3-3 zeta/delta was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-14-3-3 zeta/delta Antibody (MBS1750580) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Figure 6. IHC analysis of 14-3-3 zeta/delta using anti-14-3-3 zeta/delta antibody (MBS1750580).
14-3-3 zeta/delta was detected in paraffin-embedded section of rat kidney tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-14-3-3 zeta/delta Antibody (MBS1750580) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.