Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Form:
Lyophilized
Target:
Galactose-1-phosphate uridylyltransferase
Application Dilute:
Western blot, 0.1-0.5µg/ml Immunohistochemistry (Paraffin-embedded Section), 2-5µg/ml Immunoprecipitation, 0.5-2 µg/ml, Human Flow Cytometry(Fixed), 1-3 µg/1x10 6 cells ELISA, 0.1-0.5µg/ml
WB analysis of GALT using anti-GALT antibody.Lane 1:human HepG2 Cells,2:human MCF-7 Cells.
Flow Cytometry analysis of HepG2 cells using anti-GALT antibody. Overlay histogram showing HepG2 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GALT Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
IHC analysis of GALT using anti-GALT antibody. GALT was detected in a 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 2 µg/ml rabbit anti-GALT Antibody overnight at 4C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Immunoprecipitating GALT in THP-1 whole cell lysate. Western blot analysis of GALT using anti-GALT antibody, Lane 1: THP-1 whole cell lysates (30 ug), Lane 2: Rabbit control IgG instead of anti-GALT antibody in THP-1 whole cell lysate, Lane 3: anti-GALT antibody (2 µg) + THP-1 whole cell lysate (500 µg). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-GALT antigen affinity purified polyclonal antibody at a dilution of 0.5 µg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. A specific band was detected for GALT at approximately 43 kDa. The expected band size for GALT is at 43 kDa.
Western blot analysis of GALT using anti-GALT antibody. 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 30 ug of sample under reducing conditions. Lane 1: human 293T whole cell lysates, Lane 2: human THP-1 whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human Hela whole cell lysates, Lane 5: rat lung tissue lysates, Lane 6: rat kidney tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-GALT antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4C, 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 1:5000 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 GALT at approximately 43 kDa. The expected band size for GALT is at 43 kDa.
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