Anti-SIRT1 Antibody Picoband®

Sirtuin 6 is the possible contributor to gender differences upon IRI. A Graphical representations show the relative abundance of Sirtuin 1-7 mRNA in different groups. ** P < 0.01, *** P < 0.001 versus sham controls in male group ( n = 5); † P < 0.05 versus sham controls in female group ( n = 5). B , C Representative western blot ( B ) and graphical representations of ( C ) Sirtuin 1-7 protein expression levels are shown. * P < 0.05, ** P < 0.01, *** P < 0.001 versus male group ( n = 5). D Representative micrographs showing the expression of Sirtuin 6 in different groups, as indicated. Paraffin-embedded kidney sections were stained with an antibody against Sirtuin 6. Arrows indicate positive staining. Scale bar, 50 μm. E Representative micrographs showing the expression of PGC-1α and TOMM20 in different groups, as indicated. Frozen kidney sections were stained with an antibody against PGC-1α and TOMM20. Arrows indicate positive staining. Scale bar, 50 μm. F – H Representative western blot ( F ) and graphical representations of ( G ) PGC-1α and ( H ) TOMM20 protein expression levels are shown. *** P < 0.001 versus male group ( n = 5).
Index in PubMed under a CC BY license. PMID: 37185276

The expression of Sirtuin 6 was the key regulator for gender differences in rhabdomyolysis-induced AKI. A , B Representative western blot ( A ) and graphical representations of ( B ) Sirtuin 1-7 protein expression levels are shown. * P < 0.05, ** P < 0.01, *** P < 0.001 versus female group ( n = 5). C Representative micrographs show the expression of Sirtuin 6 in different groups, as indicated. Paraffin-embedded kidney sections were stained with an antibody against Sirtuin 6. Arrows indicate positive staining. Scale bar, 50 μm. D Representative micrographs showing the expression of PGC-1α and TOMM20 in different groups, as indicated. Frozen kidney sections were stained with an antibody against PGC-1α and TOMM20. Arrows indicate positive staining. Scale bar, 50 μm. E – G Representative western blot ( E ) and graphical representations of ( F ) PGC-1α and ( G ) TOMM20 protein expression levels are shown. * P < 0.05, *** P < 0.001 versus male group ( n = 5).
Index in PubMed under a CC BY license. PMID: 37185276

AR increases acetylation of PGC-1α by downregulating Sirtuin 6 expression. A – C Representative western blot ( A ) and graphical representations of ( B ) Sirtuin 1-7 and ( C ) PGC-1α protein expression levels are shown. * P < 0.05, ** P < 0.01 versus control group ( n = 3). HKC-8 cells were transfected with pcDNA3 or AR overexpression plasmid for 24 h. D Representative ChIP assay results showing the binding of AR to the Sirtuin 6 gene promoter region. HKC‐8 cells were incubated with DHT (10 μMol/L) or not for 24 h. Cell lysates were precipitated with an antibody against AR, histone H3, or nonimmune IgG, and the ChIP assay was performed for Sirtuin 6 gene promoters. Total diluted lysate was used as the total genomic input DNA. E Graphical representations show the relative abundance of Sirtuin 1-7 mRNA in different groups. ** P < 0.01 versus control group ( n = 3). F – I Representative western blot ( F ) and graphical representations of ( G ) Sirtuin 1-7, ( H ) PGC-1α and ( I ) AR protein expression levels are shown. HKC-8 cells were incubated with DHT (10 μMol/L) and transfected with AR-shRNA for 24 h. * P < 0.05, ** P < 0.01, *** P < 0.001 ( n = 3); † P < 0.05, †† P < 0.01, ††† P < 0.001 ( n = 3). J Representative graphs show the binding of PGC-1α with Sirtuin 6 or acetyl. HKC-8 cells were transfected with pcDNA3 or AR overexpression plasmid for 24 h. K Representative graphs show the binding of PGC-1α with acetyl, and the expression of PGC-1α in different groups, as indicated. HKC-8 cells were treated with DHT (10 μMol/L) and transfected with Sirtuin 6 overexpression plasmid for 24 h. L Representative graphs show the binding of PGC-1α with acetyl, and the protein levels of AR in nuclear fractions in different groups, as indicated. HKC-8 cells were treated with DHT (10 μMol/L) for 24 h. M Representative graphs show the binding of PGC-1α with acetyl, and the protein levels of AR in nuclear fractions in sham control and IRI group in male mice, as indicated.
Index in PubMed under a CC BY license. PMID: 37185276

Western blot analysis of SIRT1 using anti-SIRT1 antibody (PB9159).
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 under reducing conditions.
Lane 1: human HepG2 whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: huamn SiHa whole cell lysates,
Lane 4: huamn Daudi whole cell lysates,
Lane 5: huamn K562 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-SIRT1 antigen affinity purified polyclonal antibody (Catalog # PB9159) 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 SIRT1 at approximately 120 kDa. The expected band size for SIRT1 is at 82 kDa.

IF analysis of SIRT1 using anti-SIRT1 antibody (PB9159).
SIRT1 was detected in an immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-SIRT1 Antibody (PB9159) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 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.