timp 1 antibody  (Boster Bio)

Journal: Clinical Ophthalmology (Auckland, N.Z.)

Article Title: Sorsby Fundus Dystrophy in an Asian Pedigree: Pathogenic Timp3 P.Y191c Variant Impairs Its Binding with Mmp2/9 and Cellular Localization

doi: 10.2147/OPTH.S556592

Figure Lengend Snippet: Whole exome sequencing. ( A ) Sanger sequencing chromatograms of twelve family members identified the heterozygous c.572A>G (p. Y191C ) TIMP3 variant in the proband (III:5), his father (II:6), uncle (II:3, II:8), aunt (II:11), and cousins (III:3, III:10). ( B ) Pedigree analysis illustrating the inheritance of the c.572A>G (p. Y191C ) variant. ( C ) Multiple sequence alignments of TIMP3 Tyr191 across species revealed that the variant occurred in highly conserved residues. ( D ) Schematic structures of the original and mutant amino acids, highlighting the backbone in red and the side chain in black. ( E ) Comparative 3D structures of the wild-type ( a ) and mutant ( b ) proteins.

Article Snippet: Primary antibodies included TIMP3 (Proteintech, 10858-1-AP, WB: 1:2000, IP: 1:1000, IF: 1:400), MMP9 (Proteintech, 10375-2-AP, WB: 1:600, IP:1:500, IF: 1:100), MMP2 (Proteintech, 66366-1-Ig, WB: 1:1000, IP:1:500, IF: 1:200), and GAPDH (Abcam, ab8245, 1:5000).

Techniques: Sequencing, Variant Assay, Mutagenesis

Journal: Clinical Ophthalmology (Auckland, N.Z.)

Article Title: Sorsby Fundus Dystrophy in an Asian Pedigree: Pathogenic Timp3 P.Y191c Variant Impairs Its Binding with Mmp2/9 and Cellular Localization

doi: 10.2147/OPTH.S556592

Figure Lengend Snippet: The TIMP3 variant Y191C mitigated the cell viability and promoted the apoptosis of ARPE-19. ( A ) CCK-8 assay demonstrated a significant reduction in cell viability in the MT group compared to the empty vector control group 24 hours post-LPS treatment ( P= 0.0096), while no significant difference was observed in the WT group ( P= 0.7128). ( B and C ) Flow cytometry analysis indicated a significant increase in apoptosis rates in the MT group compared to the empty vector control group following LPS treatment ( P= 0.0005). In contrast, the WT group exhibited a 10% reduction in apoptosis rates compared to the empty vector control group. FIGURE 4 The TIMP3 variant Y191C mitigated the cell viability and promoted the apoptosis of ARPE-19. ( A ) CCK-8 assay demonstrated a significant reduction in cell viability in the MT group compared to the empty vector control group 24 hours post-LPS treatment ( P= 0.0096), while no significant difference was observed in the WT group ( P= 0.7128). ( B and C ) Flow cytometry analysis indicated a significant increase in apoptosis rates in the MT group compared to the empty vector control group following LPS treatment ( P= 0.0005). In contrast, the WT group exhibited a 10% reduction in apoptosis rates compared to the empty vector control group. n=3/group. “n” denotes biological replicates, defined as independently assayed aliquots derived from the same lentiviral infection and differentiation batch.

Article Snippet: Primary antibodies included TIMP3 (Proteintech, 10858-1-AP, WB: 1:2000, IP: 1:1000, IF: 1:400), MMP9 (Proteintech, 10375-2-AP, WB: 1:600, IP:1:500, IF: 1:100), MMP2 (Proteintech, 66366-1-Ig, WB: 1:1000, IP:1:500, IF: 1:200), and GAPDH (Abcam, ab8245, 1:5000).

Techniques: Variant Assay, CCK-8 Assay, Plasmid Preparation, Control, Flow Cytometry, Derivative Assay, Infection

Journal: Clinical Ophthalmology (Auckland, N.Z.)

Article Title: Sorsby Fundus Dystrophy in an Asian Pedigree: Pathogenic Timp3 P.Y191c Variant Impairs Its Binding with Mmp2/9 and Cellular Localization

doi: 10.2147/OPTH.S556592

Figure Lengend Snippet: The Y191C variant inhibited the interaction between TIMP3 and MMP proteins. ( A and B ) The Y191C mutation increases FLAG-TIMP3 complex formation, selectively enhancing interaction with MMP9 proteolytic fragments (45~55 kDa) while reducing MMP2 binding. ( C – E ) Following LPS administration, MMP2 expression levels in the empty vector control group remained comparable to those in the normal control group. However, in the MT group, MMP2 expression was significantly elevated compared to the empty vector group ( P < 0.0001, 1.84 ± 0.21-fold) and showed a modest increase compared to the WT control group ( P= 0.0889, 1.15 ± 0.13-fold). Additionally, MMP9 expression in the MT group was markedly higher than in the empty vector group after LPS treatment ( P < 0.0172, 2.12 ± 0.41-fold). ( F ) IF staining revealed nuclear localization of MMP2 in ARPE-19 cells of the WT group, whereas in the MT group, MMP2 expression extended into the cytoplasm of ARPE-19 cells. n=3/group. “n” refers to the number of independent biological replicates.

Article Snippet: Primary antibodies included TIMP3 (Proteintech, 10858-1-AP, WB: 1:2000, IP: 1:1000, IF: 1:400), MMP9 (Proteintech, 10375-2-AP, WB: 1:600, IP:1:500, IF: 1:100), MMP2 (Proteintech, 66366-1-Ig, WB: 1:1000, IP:1:500, IF: 1:200), and GAPDH (Abcam, ab8245, 1:5000).

Techniques: Variant Assay, Mutagenesis, Binding Assay, Expressing, Plasmid Preparation, Control, Staining

Journal: Journal of Inflammation Research

Article Title: TIMP4 as a Potential Complementary Biomarker and Therapeutic Target in Membranous Nephropathy: A Multi-Omics Investigation with Clinical Validation

doi: 10.2147/JIR.S545422

Figure Lengend Snippet: The results of Bayesian colocalization analysis. ( A ) TIMP4; ( B ) LY75. The figure illustrates meaningful colocalization results based on plasma proteins, with the instrumental variables and their chromosomal positions visualized.

Article Snippet: TIMP4 concentrations were quantified using a commercial enzyme-linked immunosorbent assay (ELISA) system (KE00198; Proteintech) according to the manufacturer’s protocols.

Techniques: Clinical Proteomics

Journal: Journal of Inflammation Research

Article Title: TIMP4 as a Potential Complementary Biomarker and Therapeutic Target in Membranous Nephropathy: A Multi-Omics Investigation with Clinical Validation

doi: 10.2147/JIR.S545422

Figure Lengend Snippet: Validation of TIMP4 protein expression. ( A ) TIMP4 levels in MN patients vs controls. ( B ) TIMP4 expression across PLA2R-positive MN, PLA2R-negative MN, and control groups. *** P < 0.001; ns, not significant ( P > 0.05).

Article Snippet: TIMP4 concentrations were quantified using a commercial enzyme-linked immunosorbent assay (ELISA) system (KE00198; Proteintech) according to the manufacturer’s protocols.

Techniques: Biomarker Discovery, Expressing, Control

Journal: Journal of Inflammation Research

Article Title: TIMP4 as a Potential Complementary Biomarker and Therapeutic Target in Membranous Nephropathy: A Multi-Omics Investigation with Clinical Validation

doi: 10.2147/JIR.S545422

Figure Lengend Snippet: Single-cell RNA sequencing profile of kidney tissues. ( A ) UMAP projection after Harmony batch correction, integrating datasets across samples. ( B ) Eighteen distinct clusters visualized by UMAP plotting. ( C ) Dot plot illustrating the expression of marker genes in different cell types. ( D ) Annotation of 13 major cell types based on marker genes. ( E ) Violin plot depicting TIMP4 expression levels across different cell populations.

Article Snippet: TIMP4 concentrations were quantified using a commercial enzyme-linked immunosorbent assay (ELISA) system (KE00198; Proteintech) according to the manufacturer’s protocols.

Techniques: RNA Sequencing, Expressing, Marker