Western blot (WB): | 1:500-2000 |
Immunohistochemistry (IHC): | 1:50-400 |
Immunocytochemistry/Immunofluorescence (ICC/IF): | 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 SLC2A1 using anti-SLC2A1 antibody (PB9435). The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human placenta tissue lysate, Lane 2: human A431 whole cell lysates,Lane 3: mouse brain tissue lysate, Lane 4: rat PC-12 whole cell lysates,Lane 5: mouse NIH3T3 whole cell lysate. 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 rabbit anti-SLC2A1 antigen affinity purified polyclonal antibody (Catalog # PB9435) at 0.5 μg/mL overnight at 4°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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SLC2A1 at approximately 55KD. The expected band size for SLC2A1 is at 55KD.
Figure 2. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of Mouse Brain 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-SLC2A1 Antibody (PB9435) 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. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of Rat Brain 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-SLC2A1 Antibody (PB9435) 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 4. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of Rat Brain 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-SLC2A1 Antibody (PB9435) 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 5. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of Human Placenta 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-SLC2A1 Antibody (PB9435) 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 6. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of Human Placenta 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-SLC2A1 Antibody (PB9435) 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 7. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of human mammary 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-SLC2A1 Antibody (PB9435) 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 8. ICC analysis of anti- SLC2A1 antibody (PB9435). Was detected in immunocytochemical section of HepG2 cells. Cells were stained using the Dylight488-conjugated Anti-rabbit IgG Secondary Antibody (green)(Catalog # BA1127) and counterstained with DAPI (blue).
Figure 9. Flow cytometry analysis of A431 cell(1:100) DyLight488 conjugated goat anti-rabbit IgG(blue) was used as secondary antibody. Isotype control antibody (Green line) was rabbit IgG DyLight488. Unlabelled sample (Red line).
Figure 1. Western blot analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435). The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human placenta tissue lysate, Lane 2: human A431 whole cell lysates,Lane 3: mouse brain tissue lysate, Lane 4: rat PC-12 whole cell lysates,Lane 5: mouse NIH3T3 whole cell lysate. 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 rabbit anti-SLC2A1 antigen affinity purified polyclonal antibody (Catalog # PB9435) at 0.5 μg/mL overnight at 4°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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SLC2A1 at approximately 55KD. The expected band size for SLC2A1 is at 55KD.
Figure 2. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of Mouse Brain 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-SLC2A1 Antibody (PB9435) 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. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of Rat Brain 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-SLC2A1 Antibody (PB9435) 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 4. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of Rat Brain 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-SLC2A1 Antibody (PB9435) 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 5. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of Human Placenta 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-SLC2A1 Antibody (PB9435) 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 6. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of Human Placenta 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-SLC2A1 Antibody (PB9435) 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 7. IHC analysis of SLC2A1 using anti-SLC2A1 antibody (PB9435).SLC2A1 was detected in paraffin-embedded section of human mammary 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-SLC2A1 Antibody (PB9435) 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 8. ICC analysis of anti- SLC2A1 antibody (PB9435). Was detected in immunocytochemical section of HepG2 cells. Cells were stained using the Dylight488-conjugated Anti-rabbit IgG Secondary Antibody (green)(Catalog # BA1127) and counterstained with DAPI (blue).
Figure 9. Flow cytometry analysis of A431 cell(1:100) DyLight488 conjugated goat anti-rabbit IgG(blue) was used as secondary antibody. Isotype control antibody (Green line) was rabbit IgG DyLight488. Unlabelled sample (Red line).