Mouse ASS1 Monoclonal Antibody | anti-ASS1 antibody
Anti-ASS1 Antibody (monoclonal, 5I5)
IHC-P: 2-5ug/ml|Human, Mouse, Rat|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
FCM (Flow Cytometry)
(Figure 8. Flow Cytometry analysis of SiHa cells using anti- ASS1 antibody (AAA19369).Overlay histogram showing SiHa cells stained with AAA19369 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ASS1 Antibody (AAA19369, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)
IF (Immunofluorescence)
(Figure 7. IF analysis of ASS1 using anti- ASS1 antibody (AAA19369).ASS1 was detected in immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- ASS1 Antibody (AAA19369) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) 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.)
IHC (Immunohistchemistry)
(Figure 6. IHC analysis of ASS1 using anti-ASS1 antibody (AAA19369).ASS1 was detected in paraffin-embedded section of mouse kidney 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 2μg/ml mouse anti-ASS1 Antibody (AAA19369) overnight at 4 degree C. Biotinylated goat anti-mouse 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 # with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 5. IHC analysis of ASS1 using anti-ASS1 antibody (AAA19369).ASS1 was detected in paraffin-embedded section of rat liver 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 2μg/ml mouse anti-ASS1 Antibody (AAA19369) overnight at 4 degree C. Biotinylated goat anti-mouse 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 # with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 4. IHC analysis of ASS1 using anti-ASS1 antibody (AAA19369).ASS1 was detected in paraffin-embedded section of human colon 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 2μg/ml mouse anti-ASS1 Antibody (AAA19369) overnight at 4 degree C. Biotinylated goat anti-mouse 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 # with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 3. IHC analysis of ASS1 using anti-ASS1 antibody (AAA19369).ASS1 was detected in paraffin-embedded section of human breast 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 2μg/ml mouse anti-ASS1 Antibody (AAA19369) overnight at 4 degree C. Biotinylated goat anti-mouse 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 # with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 2. IHC analysis of ASS1 using anti-ASS1 antibody (AAA19369).ASS1 was detected in paraffin-embedded section of human prostate 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 2μg/ml mouse anti-ASS1 Antibody (AAA19369) overnight at 4 degree C. Biotinylated goat anti-mouse 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 # with DAB as the chromogen.)
WB (Western Blot)
(Figure 1. Western blot analysis of ASS1 using anti-ASS1 antibody (AAA19369).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 lysatesLane 2: human HEPG2 whole cell lysatesLane 3: human Hek293 wohle cell lysatesLane 4: monkey kidney tissue lysatesLane 5: rat liver tissue lysatesLane 6: rat kidney tissue lysatesLane 7: mouse liver tissue lysatesLane 8: 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 mouse anti-ASS1 antigen affinity purified monoclonal antibody (Catalog # AAA19369) at 0. 5 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-mouse 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 # with Tanon 5200 system. A specific band was detected for ASS1 at approximately 47KD. The expected band size for ASS1 is at 47KD.)
2. Rabinovich, S., Adler, L., Yizhak, K., Sarver, A., Silberman, A., Agron, S., Stettner, N., Sun, Q., Brandis, A., Helbling, D., Korman, S., Itzkovitz, S., Dimmock, D., Ulitsky, I., Nagamani, S. C. S., Ruppin, E., Erez, A. Diversion of aspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis. Nature 527: 379-383, 2015.
3. Sase, M., Kobayashi, K., Imamura, Y., Saheki, T., Nakano, K., Miura, S., Mori, M. Level of translatable messenger RNA coding for argininosuccinate synthetase in the liver of the patients with quantitative-type citrullinemia. Hum. Genet. 69: 130-134, 1985.
