Anti-SLC40A1 Antibody Picoband®

Western blot analysis of SLC40A1 using anti-SLC40A1 antibody (A01953-2).
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 unde
r reducing conditions.
Lane 1: human placenta tissue lysates,
Lane 2: rat liver tissue lysates,
Lane 3: mouse spleen tissue lysates,
Lane 4: mouse liver 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-SLC40A1 antigen affinity purified polyclonal antibody (Catalog # A01953-2) 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 1:5000 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 SLC40A1 at approximately 70 kDa. The expected band size for SLC40A1 is at 63 kDa.

FSGS rats showed iron metabolism disorder. ( A ) Prussian blue staining (scale bar: 100 μm). ( B ) Fe 2+ results. ( C ) Western blot analysis of hepcidin, ferroportin and TFR. ( D ) Relative expression of hepcidin. ( E ) Relative expression of ferroportin. ( F ) Relative expression of TFR. * P < 0.05.
Index in PubMed under a CC BY license. PMID: 38097813

IHC analysis of SLC40A1 using anti-SLC40A1 antibody (A01953-2).
SLC40A1 was detected in a paraffin-embedded section of human larynx squamous cell carcinoma 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 2 μg/ml rabbit anti-SLC40A1 Antibody (A01953-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of SLC40A1 using anti-SLC40A1 antibody (A01953-2).
SLC40A1 was detected in a paraffin-embedded section of human liver 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 2 μg/ml rabbit anti-SLC40A1 Antibody (A01953-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of SLC40A1 using anti-SLC40A1 antibody (A01953-2).
SLC40A1 was detected in a paraffin-embedded section of human ovarian serous adenocarcinoma 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 2 μg/ml rabbit anti-SLC40A1 Antibody (A01953-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of SLC40A1 using anti-SLC40A1 antibody (A01953-2).
SLC40A1 was detected in a paraffin-embedded section of human prostate adenocarcinoma 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 2 μg/ml rabbit anti-SLC40A1 Antibody (A01953-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of SLC40A1 using anti-SLC40A1 antibody (A01953-2).
SLC40A1 was detected in a paraffin-embedded section of human rectum adenocarcinoma 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 2 μg/ml rabbit anti-SLC40A1 Antibody (A01953-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of SLC40A1 using anti-SLC40A1 antibody (A01953-2).
SLC40A1 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 5 μg/mL rabbit anti-SLC40A1 Antibody (A01953-2) overnight at 4°C. FITC Conjugated Goat Anti-Rabbit IgG (BA1105) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of U20S cells using anti-SLC40A1 antibody (A01953-2).
Overlay histogram showing U20S cells stained with A01953-2 (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-SLC40A1 Antibody (A01953-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of SLC40A1 using anti-SLC40A1 antibody (A04887-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1-5: model group-human uterine tissue lysates,
Lane 6-10: young group-human uterine 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-SLC40A1 antigen affinity purified polyclonal antibody (A04887-1) at 1:1000 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 1:5000 for 1 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate with Tanon 5200 system. A specific band was detected for SLC40A1 at approximately 60-70 kDa. The expected band size for SLC40A1 is at 70 kDa.