Anti-TIM 1/HAVCR1 Antibody Picoband®

Western blot analysis of HAVCR1 using anti-HAVCR1 antibody (PA1632).
Electrophoresis was performed on a 10% 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: rat kidney 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-HAVCR1 antigen affinity purified polyclonal antibody (PA1632) 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 (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for HAVCR1 at approximately 50 kDa. The expected band size for HAVCR1 is at 34 kDa.

IHC analysis of HAVCR1 using anti-HAVCR1 antibody (PA1632).
HAVCR1 was detected in a paraffin-embedded section of rat kidney 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-HAVCR1 Antibody (PA1632) 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.

The ectopic expression of Sirtuin 6 relieves renal injury and mitochondrial dysfunction upon IRI. A Experimental design. Green arrow showed the injection of pcDNA plasmid or pFlag-Sirtuin 6 overexpression plasmid. Male mice were subjected to IRI or sham respectively, and euthanized 24 h after IRI. B Scr levels in three groups, as indicated. Scr was expressed as milligrams per deciliter. *** P < 0.001 versus sham controls ( n = 5); †† P < 0.01 versus pcDNA group ( n = 5). C Representative western blot of flag tag is shown. 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 versus sham controls ( n = 5); †† P < 0.01, ††† P < 0.001 versus pcDNA group ( n = 5). H Representative micrographs show renal tubular morphologic injury, the expression of KIM-1 and caspase-3, and TUNEL assay in different groups, as indicated. Paraffin sections were subjected to periodic acid–Schiff (PAS) staining, stained with an antibody against KIM-1 and caspase-3. Frozen kidney sections were subjected to TUNEL staining. Arrows indicate positive staining. Scale bar, 50 μm. I – M Representative western blot ( I ) and graphical representations of ( J ) KIM-1, ( K ) FAS-L, ( L ) Bax and ( M ) cleaved caspase-3 protein expression levels are shown. * P < 0.05, ** P < 0.01, *** P < 0.001 versus sham controls ( n = 5); † P < 0.05, †† P < 0.01 versus pcDNA group ( n = 5). N Tubular injury score depending on PAS staining in three groups, as indicated. *** P < 0.001 versus sham controls ( n = 5); ††† P < 0.001 versus pcDNA group ( n = 5).
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The ectopic knockdown of AR ameliorates renal injury and mitochondrial dysfunction upon IRI. A Experimental design. Green arrow showed the injection of control-shRNA (pLVX-shRNA) or AR-shRNA (pLVX-shAR) plasmid. Male mice were subjected to IRI or sham respectively, and euthanized 24 h after IRI. B Scr levels in three groups, as indicated. Scr was expressed as milligrams per deciliter. ** P < 0.01 versus sham controls ( n = 5); †† P < 0.01 versus control-shRNA group ( n = 5). C BUN levels in three groups, as indicated. BUN was expressed as milligrams per deciliter. ** P < 0.01 versus sham controls ( n = 5); † P < 0.05 versus control-shRNA group ( n = 5). D , E Representative western blot ( D ) and graphical representations of ( E ) AR protein expression levels are shown. *** P < 0.001 versus sham controls ( n = 5); †† P < 0.01 versus control-shRNA group ( n = 5). F , G Representative western blot ( F ) and graphical representations of ( G ) Sirtuin 6 protein expression levels are shown. ** P < 0.01 versus sham controls ( n = 5); †† P < 0.01 versus control-shRNA group ( n = 5). H Representative micrographs showing the expression of Sirtuin 6 in different groups. Paraffin sections were stained with an antibody against Sirtuin 6. Arrows indicate positive staining. Scale bar, 50 μm. I Representative micrographs show renal tubular morphologic injury, the expression of KIM-1 and caspase-3, and TUNEL assay in different groups, as indicated. Paraffin sections were subjected to PAS staining, stained with an antibody against KIM-1 and caspase-3. Frozen kidney sections were subjected to TUNEL staining. Arrows indicate positive staining. Scale bar, 50 μm. (J – N) Representative western blot ( J ) and graphical representations of ( K ) KIM-1, ( L ) FAS-L, ( M ) Bax and ( N ) cleaved caspase-3 protein expression levels are shown. ** P < 0.01, *** P < 0.001 versus sham controls ( n = 5); †† P < 0.01, ††† P < 0.001 versus control-shRNA group ( n = 5). O Tubular injury scores depending on PAS staining in three groups, as indicated. *** P < 0.001 versus sham controls ( n = 5); ††† P < 0.001 versus control-shRNA group ( n = 5). P 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. Q – S Representative western blot ( Q ) and graphical representations of ( R ) PGC-1α and ( S ) TOMM20 protein expression levels are shown. *** P < 0.001 versus sham controls ( n = 5); ††† P < 0.001 versus control-shRNA group ( n = 5).
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Male mice were more susceptive to rhabdomyolysis-induced AKI and tubular apoptosis in kidney. A Experimental design. Female and male mice were intramuscularly injected with 50% glycerol at the dose of 7.5 ml/kg or normal saline respectively. Mice were euthanized 3 days after intramuscular injection. B Scr levels in four groups, as indicated. Scr was expressed as milligrams per deciliter. ** P < 0.01 versus sham controls in male group ( n = 5); †† P < 0.01 versus sham controls in female group ( n = 5); # P < 0.05 versus male group in glycerol group ( n = 5). C Graphical representations show three day-mortality in different genders after glycerol administration, as indicated. * P < 0.05 versus male group (n of male group = 20; n of female group = 17). D Representative micrographs show renal tubular morphologic injury and the expression of KIM-1 in different groups, as indicated. Paraffin sections were subjected to PAS staining and stained with an antibody against KIM-1. Arrows indicate positive staining. Scale bar, 50 μm. E Tubular injury score depending on PAS staining in four groups, as indicated. *** P < 0.001 versus sham controls in male group ( n = 5); ††† P < 0.001 versus sham controls in female group ( n = 5). F – J Representative western blot ( F ) and graphical representations of ( G ) KIM-1, ( H ) FAS-L, ( I ) Bax and ( J ) cleaved caspase-3 protein expression levels are shown. * P < 0.05, ** P < 0.01, *** P < 0.001 versus female group ( n = 5). K Representative micrographs show the expression of caspase-3 and TUNEL staining in different groups, as indicated. Paraffin sections were stained with an antibody against caspase-3. Frozen kidney sections were subjected to TUNEL staining. Arrow indicates positive staining. Scale bar, 50 μm.
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Ectopic CD44 aggravates tubular cell apoptosis and kidney injury in IRI mice. A Experimental design: Green arrow indicated the injection of pcDNA3 plasmid or p-HA-CD44 overexpression plasmid. Mice were subjected to IRI surgery or sham surgery, respectively, as shown in the red arrow. Mice are euthanized 24 h after surgery. B Scr levels in three groups, as indicated. Scr was expressed as milligrams per deciliter. * P < 0.05 versus sham group; # P < 0.05 versus IRI group injected with pcDNA3 ( n = 5). C BUN levels in three groups, as indicated. BUN was expressed as milligrams per deciliter. ** P < 0.01 versus sham group; # P < 0.05 versus IRI group injected with pcDNA3 ( n = 5). D and E Representative western blot of CD44 ( D ) and graphical presentations ( E ) of protein levels are shown. ** P < 0.01 versus sham group; # P < 0.05 versus IRI group injected with pcDNA3 ( n = 5). F Representative micrographs show the expression of CD44, and TUNEL assay in different groups, as indicated. Arrows indicate positive staining. Frozen kidney sections were subjected to TUNEL assay or stained with an antibody against CD44. Scale bar, 50 μm. G–J Representative western blot ( G ) and graphical presentations of H BCL2, I BAX, and J cleaved caspase 3 protein expression levels are shown. * P < 0.05, ** P < 0.01 versus sham group; # P < 0.05, ### P < 0.001 versus IRI group injected with pcDNA3 ( n = 5). K and L Representative western blot of NGAL ( K ) and graphical presentations ( L ) of protein levels are shown. ** P < 0.01 versus sham group; ## P < 0.01 versus IRI group injected with pcDNA3 ( n = 5). M Representative micrographs show renal tubular morphologic injury and the expression of KIM-1 in different groups, as indicated. Paraffin sections were subjected to PAS staining, and were stained with an antibody against KIM-1. Arrows indicate positive staining. Scale bar, 50 μm.
Index in PubMed under a CC BY license. PMID: 39979265

Male mice were more susceptive to IRI and tubular apoptosis in kidney. A Experimental design. Female and male mice were subjected to IRI or sham respectively, and euthanized 24 h after IRI. B Scr levels in four groups, as indicated. Scr was expressed as milligrams per deciliter. ** P < 0.01 versus sham controls in male group ( n = 5). C BUN levels in four groups, as indicated. BUN was expressed as milligrams per deciliter. * P < 0.05 versus sham controls in male group ( n = 5). D Representative micrographs show renal tubular morphologic injury, the expression of KIM-1 and caspase-3, and TUNEL assay in different groups, as indicated. Paraffin sections were subjected to periodic acid–Schiff (PAS) staining, stained with an antibody against KIM-1 and caspase-3. Frozen kidney sections were subjected to TUNEL assay. Arrows indicate positive staining. Scale bar, 50 μm. E – I Representative western blot ( E ) and graphical representations of ( F ) FAS-L, ( G ) Bax, ( H ) cleaved caspase-3 and ( I ) KIM-1 protein expression levels are shown. * P < 0.05, ** P < 0.01 versus male group ( n = 5).
Index in PubMed under a CC BY license. PMID: 37185276