Glutaminase/GLS (GLS) Antibody – Rabbit Polyclonal

Figure 1. Western blot analysis of Glutaminase/GLS using anti-Glutaminase/GLS antibody (MBS1753422).
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 placenta tissue lysates
Lane 2: human U87 whole cell lysates
Lane 3: human Hela whole cell lysates
Lane 4: monkey kidney tissue lysates
Lane 5: rat brain tissue lysates
Lane 6: rat kidney tissue lysates
Lane 7: rat heart tissue lysates.
Lane 8: rat skeletal muscle tissue lysates.
Lane 9: mouse brain tissue lysates.
Lane 10: mouse kidney tissue lysates.
Lane 11: mouse heart tissue lysates.
Lane 12: mouse skeletal muscle tissue lysates.
Lane 13: mouse Neuro-2a 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-Glutaminase/GLS antigen affinity purified polyclonal antibody (Catalog # MBS1753422) at 0. 25 μg/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 (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for Glutaminase/GLS at approximately 65-73KD. The expected band size for Glutaminase/GLS is at 73KD.

Figure 2. IHC analysis of Glutaminase/GLS using anti-Glutaminase/GLS antibody (MBS1753422).
Glutaminase/GLS was detected in paraffin-embedded section of human gastric cancer 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 1μg/ml rabbit anti-Glutaminase/GLS Antibody (MBS1753422) 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) (Catalog # MBS176451) with DAB as the chromogen.

Figure 3. IHC analysis of Glutaminase/GLS using anti-Glutaminase/GLS antibody (MBS1753422).
Glutaminase/GLS was detected in paraffin-embedded section of rat brain 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 1μg/ml rabbit anti-Glutaminase/GLS Antibody (MBS1753422) 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) (Catalog # MBS176451) with DAB as the chromogen.

Figure 4. IF analysis of Glutaminase/GLS using anti-Glutaminase/GLS antibody (MBS1753422).
Glutaminase/GLS was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 4μg/mL rabbit anti-Glutaminase/GLS Antibody (MBS1753422) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 5. Flow Cytometry analysis of 293T cells using anti-Glutaminase/GLS antibody (MBS1753422).
Overlay histogram showing 293T cells stained with MBS1753422 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Glutaminase/GLS Antibody (MBS1753422,1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 6. Flow Cytometry analysis of SiHa cells using anti-Glutaminase/GLS antibody (MBS1753422).
Overlay histogram showing SiHa cells stained with MBS1753422 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Glutaminase/GLS Antibody (MBS1753422,1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.