Anti-SOAT1 Antibody Picoband®

Western blot analysis of SOAT1 using anti-SOAT1 antibody (A05096-1).
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 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HUH-7 whole cell lysates,
Lane 3: human THP-1 whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: 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-SOAT1 antigen affinity purified polyclonal antibody (Catalog # A05096-1) 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 1:5000 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 SOAT1 at approximately 75 kDa. The expected band size for SOAT1 is at 65 kDa.

SOAT1 promotes the malignant progression of HCC. A Up-regulated and down-regulated differentially expression genes (DEGs) in five mRNA expression profiles. B The GO category for DEGs. BP biological process, CC cellular component, MF molecular function. The color represents the P value, and the size indicates the enrichment gene number of each pathway. C KEGG enrichment pathways of DEGs. EIP Environmental Information Processing, CP Cellular Processes, OS Organismal Systems, GIP Genetic Information Processing, HD Human Diseases, M Metabolism. D Protein–protein interaction network of DEGs. E Up-regulated gene expression of lipid metabolism and tumor progression. F Representative images of IHC staining for SOAT1 of normal liver and HCC tissues cited from The Human Protein Atlas. G SOAT1 expression level in normal tissues and HCC tissues based on the TCGA dataset. H , I Analysis of the SOAT1 expression levels in TCGA HCC samples based on the individual clinical stage ( H ) and pathological grade ( I ). J High SOAT1 expression is positively correlated with poor survival ( P = 0.0175). K Representative images of positive and negative SOAT1 expression in different HCC tissues detected by IHC. L Analysis of the expression levels of SOAT1 in HCC patient liver tissues based on ES grade and MVI grade.
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SOAT1 promotes the EMT in HCC cells. A SOAT1 expression level in different HCC cell lines cited from CCLE database. B Western blot analysis of SOAT1 expression in HepG2 and PLC/PRF/5 cell lines. C Western blot analysis of EMT related markers in SOAT1 overexpressed or knocked down cells. D Immunofluorescence assay of E-cadherin and Vimentin in cells treated with SOAT1 overexpression or shRNA vectors. E Cell phenotype changes under SOAT1 overexpressed or knocked down treatment. F , G Migration ( F ) and invasion ( G ) of HepG2 cells transfected with SOAT1 or PLC/PRF/5 cells transfected with shSOAT1. H Cell proliferation under SOAT1 overexpressed or knocked down treatment.
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SOAT1 induces the EMT via regulating cholesterol metabolism. A , B Oil red O ( A ) and BODIPY 493/503 ( B ) staining of lipid droplets in SOAT1 overexpressed HepG2 cells and SOAT1 knocked down PLC/PRF/5 cells. The relative Oil red O and intensity of BODIPY493/503 were analyzed. C SOAT1 increased accumulation of cholesterol esters. D Cellular cholesterol distribution by Filipin III staining. E Western blot analysis of SOAT1, SREBP2, LDLR, ITGAV, and ITGB4 expression levels under SOAT1 overexpression or knockdown.
Index in PubMed under a CC BY license. PMID: 38724499

Nootkatone alleviates cholesterol metabolism disorder by targeting SOAT1. A Prediction docking score between small-molecule compounds and SOAT1. B Predicted interaction of nootkatone with cavity residues of SOAT1. C Cell viability of HCC cells with nootkatone treatment for 48 h. D , E Micrographs of Oil red O ( D ) and BODIPY ( E ) staining of lipid droplets in PLC/PRF/5 induced with cholesterol (200 μg/mL) for 24 h and then treated with nootkatone (150 and 300 µM) for 48 h. F Nootkatone decreased the content of cholesterol esters in different groups. G , H Expression of SOAT1 in different groups.
Index in PubMed under a CC BY license. PMID: 38724499

Nootkatone inhibits EMT of HCC by targeting SOAT1. A BODIPY493/503 staining of lipid droplets in Control, NK, SOAT1 and SOAT1 + NK groups. B Content of cholesterol esters in different groups. C Cholesterol distribution was determined by Filipin III staining. D , E Invasion ( D ) and migration ( E ) of PLC/PRF/5 cells with different treatments. F Immunofluorescence assay of E-cadherin and Vimentin of cells in different group. G Cell phenotype under nootkatone treatment with different concentration (150 and 300 µM). H Western blot analysis of SOAT1, SREBP2, LDLR, E-cadherin, Occludin, Vimentin, Twist1, N-cadherin, Snail1, Slug, and Fibronectin expression level in different groups.
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Nootkatone suppresses the oncogenic and metastatic effects of SOAT1 in vivo. A Representative images of subcutaneous tumor xenografts in Control, SOAT1, shSOAT1, NK and SOAT1 + NK groups. B Tumor volume in different groups. C WB analysis of SOAT1, SREBP2, LDLR, E-cadherin, Occludin, Vimentin, Twist1, N-cadherin, Snail1, Slug, and Fibronectin expression levels in tumor tissue of different groups. D Visible metastatic nodules on the surface of lungs in different groups.
Index in PubMed under a CC BY license. PMID: 38724499

Nootkatone suppresses tumorigenesis and development of NAFLD-HCC mice. A Schematic illustration of experimental procedure. B Representative macroscopic images of liver in Control, Model, NK-L, and NK-H groups. C AFP expression in serum and liver tissue of mice in different groups. D Body weight of mice in different groups. E , F Liver weight ( E ) and liver weight-to-body weight ratio ( F ). G Serum TC level in four groups. H Contents of hepatic free cholesterol and cholesterol esters in four different groups. I Serum ALT and AST level in different groups. J Morphological observations of the liver and liver tissue with Oil red O, H&E and Sirius red staining. The relative Oil red O and Sirius red were obtained through the Image J Pro software. K The protein expression level of SOAT1, SREBP2, LDLR, E-cadherin, Occludin, Vimentin, Twist1, N-cadherin, Snail1, Slug, and Fibronectin expression in liver tissue of mice in different groups.
Index in PubMed under a CC BY license. PMID: 38724499

IHC analysis of SOAT1 using anti-SOAT1 antibody (A05096-1).
SOAT1 was detected in a paraffin-embedded section of human breast 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-SOAT1 Antibody (A05096-1) 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 SOAT1 using anti-SOAT1 antibody (A05096-1).
SOAT1 was detected in a paraffin-embedded section of human breast 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-SOAT1 Antibody (A05096-1) 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 SOAT1 using anti-SOAT1 antibody (A05096-1).
SOAT1 was detected in a paraffin-embedded section of human prostatic 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-SOAT1 Antibody (A05096-1) 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 SOAT1 using anti-SOAT1 antibody (A05096-1).
SOAT1 was detected in a paraffin-embedded section of human prostatic 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-SOAT1 Antibody (A05096-1) 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 SOAT1 using anti-SOAT1 antibody (A05096-1).
SOAT1 was detected in a paraffin-embedded section of human prostatic 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-SOAT1 Antibody (A05096-1) 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 SOAT1 using anti-SOAT1 antibody (A05096-1).
SOAT1 was detected in a paraffin-embedded section of human prostatic 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-SOAT1 Antibody (A05096-1) 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 SOAT1 using anti-SOAT1 antibody (A05096-1).
SOAT1 was detected in a paraffin-embedded section of rat testis 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-SOAT1 Antibody (A05096-1) 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 SOAT1 using anti-SOAT1 antibody (A05096-1).
SOAT1 was detected in a paraffin-embedded section of rat testis 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-SOAT1 Antibody (A05096-1) 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 SOAT1 using anti-SOAT1 antibody (A05096-1).
SOAT1 was detected in an immunocytochemical section of PC-3 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-SOAT1 Antibody (A05096-1) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) 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 Caco-2 cells using anti-SOAT1 antibody (A05096-1).
Overlay histogram showing Caco-2 cells stained with A05096-1 (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-SOAT1 Antibody (A05096-1, 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 (Red line) was also used as a control.

Flow Cytometry analysis of THP-1 cells using anti-SOAT1 antibody (A05096-1).
Overlay histogram showing THP-1 cells stained with A05096-1 (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-SOAT1 Antibody (A05096-1, 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 (Red line) was also used as a control.