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Western blot analysis of Glutamine Synthetase on different lysates with Mouse anti-Glutamine Synthetase antibody (EM1902-39) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: HepG2 cell lysate
Lane 4: K-562 cell lysate
Lane 5: Jurkat cell lysate
Lane 6: HEK-293 cell lysate
Lane 7: SK-Br-3 cell lysate
Lane 8: Human liver tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 30 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1902-39) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
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Fluorescence multiplex immunohistochemical analysis of mouse liver (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-Desmin (ET1606-30, White), anti-HNF4α (HA721006, Red) and anti-GS (EM1902-39, Yellow) on liver. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of ET1606-30 (1/800 dilution), HA721006 (1/5,000 dilution) and EM1902-39 (1/2,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.
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Fluorescence multiplex immunohistochemical analysis of mouse liver (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-β-catenin (ET1601-5, Tangerine), anti-αSMA (ET1607-53, Yellow), anti-SOX9 (ET1611-56, Green), anti-Albumin (ET1702-55, Cyan) anti-GS (EM1902-39, Magenta) and anti-CK19 (ET1601-6, Orange) on liver. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in six rounds of staining: in the order of ET1601-5 (1/2,000 dilution), ET1607-53 (1/3,000 dilution), ET1611-56 (1/1,500 dilution), ET1702-55 (1/3,000 dilution), EM1902-39 (1/2,000 dilution) and ET1601-6 (1/3,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
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Immunohistochemical analysis of paraffin-embedded human spleen tissue with Mouse anti-Glutamine Synthetase antibody (EM1902-39) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-39) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Mouse anti-Glutamine Synthetase antibody (EM1902-39) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-39) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded rat liver tissue with Mouse anti-Glutamine Synthetase antibody (EM1902-39) at 1/500 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-39) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of Glutamine Synthetase was done on THP-1 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1902-39, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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