Western blot (WB): | 1:500-2000 |
Immunohistochemistry (IHC): | 1:50-400 |
Flow Cytometry (Fixed): | 1:50-200 |
(Boiling the paraffin sections in 10mM citrate buffer,pH6.0,or PH8.0 EDTA repair liquid for 20 mins is required for the staining of formalin/paraffin sections.) Optimal working dilutions must be determined by end user. |
Figure 1. Western blot analysis of anti- ATF1 antibody (PB9130). The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human K562 whole cell lysates.
Use rabbit anti- ATF1 1:1000, probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002). A specific band was detected for ATF1 at approximately 38KD. The expected band size for ATF1 is at 29KD.
Figure 2. IHC analysis of ATF1 using anti-ATF1 antibody (PB9130).ATF1 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ATF1 Antibody (PB9130) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 3. Flow Cytometry analysis of SiHa cells using anti-ATF1 antibody (PB9130).Overlay histogram showing SiHa cells stained with PB9130 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATF1 Antibody (PB9130, 1:100) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 1. Western blot analysis of anti- ATF1 antibody (PB9130). The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human K562 whole cell lysates.
Use rabbit anti- ATF1 1:1000, probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002). A specific band was detected for ATF1 at approximately 38KD. The expected band size for ATF1 is at 29KD.
Figure 2. IHC analysis of ATF1 using anti-ATF1 antibody (PB9130).ATF1 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ATF1 Antibody (PB9130) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 3. Flow Cytometry analysis of SiHa cells using anti-ATF1 antibody (PB9130).Overlay histogram showing SiHa cells stained with PB9130 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATF1 Antibody (PB9130, 1:100) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.