Rabbit anti-Human NT5C Polyclonal Antibody | anti-NT5C antibody
Anti-NT5C Antibody Picoband
IHC(Paraffin-embedded Section), 1-2ug/ml, Human
FC/FACS (Fixed): 1-3ug/1x106 cells, Human
ELISA: 0.1-0.5ug/ml
FCM (Flow Cytometry)
(Figure 6. Flow Cytometry analysis of U251 cells using anti-NT5C antibody (AAA19949).Overlay histogram showing U251 cells stained with AAA19949 (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-NT5C Antibody (AAA19949, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)
IHC (Immunohistochemistry)
(Figure 5. IHC analysis of NT5C using anti-NT5C antibody (AAA19949).NT5C was detected in a paraffin-embedded section of human ovarian cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NT5C Antibody (AAA19949) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NT5C Antibody (AAA19949) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NT5C Antibody (AAA19949) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-NT5C Antibody (AAA19949) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Lane 2: human U251 whole cell lysates,Lane 3: human 293T whole cell lysates,Lane 4: human HL-60 whole cell lysates,Lane 5: human HepG2 whole cell lysates,Lane 6: human A549 whole cell lysates,Lane 7: human U20S whole cell lysates,Lane 8: human SH-SY5Y whole cell 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-NT5C antigen affinity purified polyclonal antibody (#AAA19949) at 0.5ug/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 1:5000 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 NT5C at approximately 24 kDa. The expected band size for NT5C is at 24 kDa.)