Anti-SGLT2/SLC5A2 Antibody Picoband®

Western blot analysis of SLC5A2 using anti-SLC5A2 antibody (A03748-1).
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 50ug of sample under reducing conditions.
Lane 1: human HL-60 whole cell lysates,
Lane 2: human THP-1 whole cell lysates,
Lane 3: rat kidney tissue lysates,
Lane 4: rat spleen tissue lysates,
Lane 5: rat lung tissue lysates,
Lane 6: mouse kidney tissue lysates,
Lane 7: mouse spleen tissue lysates,
Lane 8: mouse lung tissue lysates,
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-SLC5A2 antigen affinity purified polyclonal antibody (Catalog # A03748-1) 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 SLC5A2 at approximately 73KD. The expected band size for SLC5A2 is at 73KD.

High-salt diet alters SGLT2 and Na+/K + -ATPase expression in renal tubules of DKD mice. (A) Expression of SGLT2 gene in renal tubules ( n = 6 per group). (B) Paraffin-embedded renal sections were stained with SGLT2, ATP1A1 and ATP1B1 antibodies (magnification, 400×, bar = 20 μm). (C) Histopathological assessment of SGLT2, ATP1A1 and ATP1B1 proteins ( n = 4 per group). All data are mean ± SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. Control group; ## p < 0.01 and ### p < 0.001 vs. DM group.
Index in PubMed under a CC BY license. PMID: 34987387

High salt alters SGLT2 and Na+/K + -ATPase expression in HG-treated HK-2. (A) Intracellular glucose concentration. (B,C) Expression of SGLT2 in HK-2 in HK-2 cultured with HG and 15 mM NaCl. (D) Expression of NKAIN4, ATP1A1, ATP1B1 and ATP1B3 genes in HK-2 after exposure to HG and different concentrations of NaCl. (E,F) Expression of ATP1A1 protein in HK-2 with HG and 15 mM NaCl cultured. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. Control group; # p < 0.05 and ## p < 0.01 vs. HG group. (G) Expression of NKAIN4 and fatty acid metabolism related genes were detected in HK-2 treated with Digoxin. * p < 0.05 and *** p < 0.001 vs. DM + DMSO group; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. HG + NaCl + DMSO group. All data are mean ± SEM, n = 3 per group.
Index in PubMed under a CC BY license. PMID: 34987387

High-salt diet attenuates gene and protein expression in diabetic mice treated with dapagliflozin. (A) The detection of ACR ( n = 6 per group). (B) Expression of α Sma, Col-1 and Slc5a2 genes in renal tubules after dapagliflozin treatment ( n = 6 per group). (C) Paraffin-embedded renal sections were stained with FN and SGLT2 antibodies (magnification, 400×, bar = 20 μm). (D) Histopathological assessment of FN and SGLT2 proteins ( n = 4 per group). Expression of genes involved in fatty acid metabolism (E) and mitochondrial biosynthesis (H) in renal tubules after dapagliflozin treatment ( n = 6 per group). (F,I) Paraffin-embedded renal sections were stained with CPT1A, ACOX1, FABP4, FASN, ATP1A1 and ATP1B1 antibodies (magnification, 400×, bar = 20 μm). (G,J) Histopathological assessment of CPT1A, ACOX1, FABP4, FASN, ATP1A1 and ATP1B1proteins ( n = 4 per group). All data are mean ± SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. DM + Saline group; # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. DM + Dapa group.
Index in PubMed under a CC BY license. PMID: 34987387

High salt attenuates gene and protein expression in HG-cultured HK-2 treated with dapagliflozin. Expression of fibrosis-related genes (A) and Na+/K + -ATPase-related genes (B) in HG-treated HK-2 after exposure to different concentrations of dapagliflozin. n = 4 per group. (C,D) Expression of SGLT2, FN and ATP1A1 proteins in HG-treated HK-2 after exposure to 5uM and 10uM dapagliflozin. n = 3 per group. (E) Expression of fatty acid metabolism-related genes. n = 3 per group. (F,G) Expression of FN, ATP1A1 and CPT1A proteins in HK-2 co-cultured with HG, 15 mM NaCl and 5 uM dapagliflozin. n = 3 per group. All data are mean ± SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. HG + DMSO group.
Index in PubMed under a CC BY license. PMID: 34987387

IHC analysis of SLC5A2 using anti-SLC5A2 antibody (A03748-1).
SLC5A2 was detected in paraffin-embedded section of human renal cancer tissues. 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-SLC5A2 Antibody (A03748-1) 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.

IHC analysis of SLC5A2 using anti-SLC5A2 antibody (A03748-1).
SLC5A2 was detected in paraffin-embedded section of mouse kidney tissues. 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-SLC5A2 Antibody (A03748-1) 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.

IHC analysis of SLC5A2 using anti-SLC5A2 antibody (A03748-1).
SLC5A2 was detected in paraffin-embedded section of rat kidney tissues. 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-SLC5A2 Antibody (A03748-1) 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.

5. Flow Cytometry analysis of HepG2 cells using anti-SLC5A2 antibody (A03748-1).
Overlay histogram showing HepG2 cells stained with A03748-1 (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-SLC5A2 Antibody (A03748-1,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Western blot analysis of SLC5A2 using anti-SLC5A2 antibody (A03748-1).
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 50ug of sample under reducing conditions.
Lane 1: human HL-60 whole cell lysates,
Lane 2: human THP-1 whole cell lysates.
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-SLC5A2 antigen affinity purified polyclonal antibody (Catalog # A03748-1) 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 SLC5A2 at approximately 73KD. The expected band size for SLC5A2 is at 73KD.

Western blot analysis of SLC5A2 using anti-SLC5A2 antibody (A03748-1).
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 50ug of sample under reducing conditions.
Lane 1: rat kidney tissue lysates,
Lane 2: rat spleen tissue lysates,
Lane 3: rat lung tissue lysates,
Lane 4: mouse kidney tissue lysates,
Lane 5: mouse spleen tissue lysates,
Lane 6: mouse lung tissue lysates.
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-SLC5A2 antigen affinity purified polyclonal antibody (Catalog # A03748-1) 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 SLC5A2 at approximately 73KD. The expected band size for SLC5A2 is at 73KD.