Mouse eRF1/ETF1 Monoclonal Antibody | anti-ETF1 antibody
Anti-eRF1/ETF1 Antibody (monoclonal, 3B6)
IHC-P: 2-5ug/ml|Human, Rat|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human, Mouse, Rat|
FCM (Flow Cytometry)
(Figure 11. Flow Cytometry analysis of RH35 cells using anti- eRF1/ETF1 antibody (AAA19382).Overlay histogram showing RH35 cells stained with AAA19382 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-eRF1/ETF1 Antibody (AAA19382, 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.)
FCM (Flow Cytometry)
(Figure 10. Flow Cytometry analysis of HEPA1-6 cells using anti- eRF1/ETF1 antibody (AAA19382).Overlay histogram showing HEPA1-6 cells stained with AAA19382 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-eRF1/ETF1 Antibody (AAA19382, 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.)
FCM (Flow Cytometry)
(Figure 9. Flow Cytometry analysis of CACO-2 cells using anti- eRF1/ETF1 antibody (AAA19382).Overlay histogram showing CACO-2 cells stained with AAA19382 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-eRF1/ETF1 Antibody (AAA19382, 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 8. IF analysis of eRF1/ETF1 using anti- eRF1/ETF1 antibody (AAA19382).eRF1/ETF1 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- eRF1/ETF1 Antibody (AAA19382) 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 (Immunohistochemistry)
(Figure 7. IHC analysis of eRF1/ETF1 using anti-eRF1/ETF1 antibody (AAA19382).eRF1/ETF1 was detected in paraffin-embedded section of human lung 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-eRF1/ETF1 Antibody (AAA19382) 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 (Immunohistchemistry)
(Figure 6. IHC analysis of eRF1/ETF1 using anti-eRF1/ETF1 antibody (AAA19382).eRF1/ETF1 was detected in paraffin-embedded section of human liver 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-eRF1/ETF1 Antibody (AAA19382) 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 eRF1/ETF1 using anti-eRF1/ETF1 antibody (AAA19382).eRF1/ETF1 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-eRF1/ETF1 Antibody (AAA19382) 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 eRF1/ETF1 using anti-eRF1/ETF1 antibody (AAA19382).eRF1/ETF1 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-eRF1/ETF1 Antibody (AAA19382) 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 eRF1/ETF1 using anti-eRF1/ETF1 antibody (AAA19382).eRF1/ETF1 was detected in paraffin-embedded section of human ovarian 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-eRF1/ETF1 Antibody (AAA19382) 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 eRF1/ETF1 using anti-eRF1/ETF1 antibody (AAA19382).eRF1/ETF1 was detected in paraffin-embedded section of human tonsil 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-eRF1/ETF1 Antibody (AAA19382) 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 eRF1/ETF1 using anti-eRF1/ETF1 antibody (AAA19382).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 Jurkat whole cell lysatesLane 3: human K562 whole cell lysatesLane 4: human Raji whole cell lysatesLane 5: human HEPG2 whole cell lysatesLane 6: rat PC-12 whole cell lysatesLane 7: mouse RAW264. 7 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 mouse anti-eRF1/ETF1 antigen affinity purified monoclonal antibody (Catalog # AAA19382) 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 eRF1/ETF1 at approximately 49KD. The expected band size for eRF1/ETF1 is at 49KD.)
2. Guenet, L., Henry, C., Toutain, B., Dubourg, C., Le Gall, J. Y., David, V., Le Treut, A. Eukaryotic translation termination factor gene (ETF1/eRF1) maps at D5S500 in a commonly deleted region of chromosome 5q31 in malignant myeloid diseases. Cytogenet. Cell Genet. 88: 82-86, 2000.
3. Hansen, L. L., Jakobsen, C. G., Justesen, J. Assignment of the human translation termination factor 1 (ETF1) to 5q31. 1 and of the proximal marker D5S1995 by radiation hybrid mapping. Cytogenet. Cell Genet. 87: 256-257, 1999.
