anti asgr1 antibodies  (Proteintech)

Journal: Investigative Ophthalmology & Visual Science

Article Title: Silencing of lncRNA HOTAIR Using Eyedrops as a Potential Treatment for Diabetes-Associated Retinal Dysregulation and Dysfunction

doi: 10.1167/iovs.67.1.24

Figure Lengend Snippet: GalNAc configurations and ASGR1 expression. ( A ) Structural diagram of GalNAc modification version 1, hereafter known as GalNAc-siHOTAIR. ( B ) Structural diagram of GalNAc modification version 2, hereafter known as GalNAc-siHOTAIR-B. ( C ) Structural diagram of GalNAc modification version 3, hereafter known as GalNAc-siHOTAIR-C. The mRNA expressions of ( D ) Asgr1 and ( E ) ASGR1 increased in response to hyperglycemia and high glucose (HG; 25 mM), respectively. ( F ) HG caused upregulation of HOTAIR compared to normal glucose (NG; 5 mM) groups, blocking ASGR1 using a neutralizing antibody (Ab) prevented knockdown of HOTAIR by GalNAc-modified siHOTAIR in HG treated cells (mRNA and lncRNA expressions represented as ratios to ACTB mRNA, and normalized to the left-most group; n = 4 for mouse Asgr1 , n = 5 for human ASGR1 , and n = 6 for HOTAIR ; data presented as mean ± SD; * = P < 0.05 as determined by Student's t -test or 1-way ANOVA followed by Tukey's HSD test).

Article Snippet: Anti-ASGR1 antibodies (5 μg/mL; Proteintech) were added to HRECs prior to GalNAc-modified siHOTAIR as a pretreatment.

Techniques: Expressing, Modification, Blocking Assay, Knockdown

Journal: Science Advances

Article Title: Targeted inhibition of hepatic de novo ceramide synthesis ameliorates MASH

doi: 10.1126/sciadv.adx2681

Figure Lengend Snippet: ( A ) Whole-body IVIS imaging of mice injected with nonlabeled LNP-siRNA (control group) or LNPs loaded Cy5-labeled siRNA. ( B ) Representative IVIS image illustrating the distribution of LNPs across various organs. ( C ) Flow cytometry analysis showing cellular uptake of Cy5-siRNA in hepatocytes. ASGR1 was used as a hepatocyte specific marker. ( D ) Distribution of Cy5-labeled siRNA among different liver cell populations in healthy mice and MASH mice fed an MCD diet for 8 weeks. Data were presented as the percentage of Cy5-positive cells within each cell type based on FACS analysis. EC, endothelial cells; HSCs, hepatic stellate cells; DCs, dendritic cells. ( E to G ) Time-course analysis of hepatic Sptlc2 mRNA levels and SPTLC2 protein levels following a single intravenous dose of LNP-siRNA (0.3 mg/kg) in MASH mice fed an MCD diet for 4 weeks. Glyceraldehyde-3-phosphate dehydrogenase was used as a loading control for Western blot analysis, and protein expression levels were normalized to that on day 2 postdosing of LNP-Scrambled siRNA (control). ( H ) Time-course analysis of hepatic Sptlc2 mRNA levels in healthy mice following siRNA treatment at 0.3 mg/kg per dose, administered either once or twice weekly. The once-weekly group received doses on days 0 and 7, while the twice-weekly group received doses on days 0, 3, 7, and 10. N = 3 mice per group. * P < 0.05, ** P < 0.01, and *** P < 0.001; one-way ANOVA.

Article Snippet: Cells were stained with an ASGR1 antibody (Proteintech, catalog no. CL488-11739, Illinois, USA) for 30 min at 4°C in the dark.

Techniques: Imaging, Injection, Control, Labeling, Flow Cytometry, Marker, Western Blot, Expressing

Journal: International journal of molecular sciences

Article Title: A High-Throughput Cell-Based Luciferase Reporter Assay for Identifying Inhibitors of ASGR1.

doi: 10.3390/ijms26104590

Figure Lengend Snippet: Figure 1. Generation and PCR identification of endogenous ASGR1-luc reporter cell line. (A) Schematic representation of the donor plasmid and the CRISPR/Cas9 targeting strategy. (B) Illustration of PCR primer amplification site. P1 and P2 represent primers WT-F and WT-R; P3 and P4 represent primers Luc-F and Luc-R. (C) PCR results of HepG2-luc-2B, HepG2-luc-3B, and HepG2-luc-B6; (D) The expected modifications of the ASGR1 locus were confirmed by Sanger sequencing (the upper base sequence represents the desired sequence, and the lower base sequence is the sequencing result of HepG2-luc-B6).

Article Snippet: After blocking with 5% non-fat milk for two hours, the NC membrane was incubated overnight at 4 ◦C with ASGR1 recombinant antibody (1:5000, Proteintech, San Diego, CA, USA).

Techniques: Plasmid Preparation, CRISPR, Amplification, Sequencing

Journal: International journal of molecular sciences

Article Title: A High-Throughput Cell-Based Luciferase Reporter Assay for Identifying Inhibitors of ASGR1.

doi: 10.3390/ijms26104590

Figure Lengend Snippet: Figure 3. The time-dependent inhibitory effects of four inhibitors on ASGR1 protein expression in HepG2 and Huh1 cells. (A–C) show HepG2 cells were harvested for immunoblotting analysis of ASGR1 expression after being treated with 10 µM concentrations of Ap-3, GlaA, OliA, and BreA for 24, 48, and 72 h. (D–F) show the similar treatment conditions in Huh1 cells.

Article Snippet: After blocking with 5% non-fat milk for two hours, the NC membrane was incubated overnight at 4 ◦C with ASGR1 recombinant antibody (1:5000, Proteintech, San Diego, CA, USA).

Techniques: Expressing, Western Blot

Journal: International journal of molecular sciences

Article Title: A High-Throughput Cell-Based Luciferase Reporter Assay for Identifying Inhibitors of ASGR1.

doi: 10.3390/ijms26104590

Figure Lengend Snippet: Figure 4. The impact of different concentrations of AP-3 on the expression levels of ASGR1 protein in HepG2 and Huh1 cell lines. (A–C) show HepG2 cells were harvested at 24 h, 48 h, and 72 h after treatment with Ap-3 at concentrations ranging from 0.1 to 20 µM for immunoblotting. (D–F) depict Huh1 cells harvested under the same treatment conditions for immunoblotting.

Article Snippet: After blocking with 5% non-fat milk for two hours, the NC membrane was incubated overnight at 4 ◦C with ASGR1 recombinant antibody (1:5000, Proteintech, San Diego, CA, USA).

Techniques: Expressing, Western Blot