67706 1 ig  (Proteintech)

Journal: Journal of Pharmaceutical Analysis

Article Title: Signatures of proteomics and glycoproteomics revealed liraglutide ameliorates MASLD by regulating specific metabolic homeostasis in mice

doi: 10.1016/j.jpha.2025.101273

Figure Lengend Snippet: Identification and validation of potential signature molecules affected by liraglutide (Lira). (A) Protein-protein interaction (PPI) analysis of differential proteins and glycoproteins in major pathways. Node size reflects degree value, with red representing glycoproteins and blue representing proteins. Color-coded segments show pathway associations. (B) Messenger RNA (mRNA) expression of Acaa2 , Lamc1 , and Col4a2 (for proteins), and Acox1 , Gclc , and Shmt2 (for glycoproteins). (C) Protein expression of laminin subunit gamma-1 (LAMC1), collagen type IV alpha-2 chain (COL4A2), and acetyl-CoA acyltransferase 2 (ACAA2) (for proteins), and glutamate-cysteine ligase catalytic subunit (GCLC) (for glycoproteins). Data is presented as mean ± standard deviation (SD) ( n = 3 per group). P values were calculated using Student's t -test or one-way analysis of variance (ANOVA) with Tukey's comparisons. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. ns: not significant. PPAR: peroxisome proliferators-activated receptor; ECM: extracellular matrix; ND: normal diet; HFD: high-fat diet; HFDL: HFD + Lira; GAPDH: glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The primary antibodies used in our study were as follows: laminin subunit gamma-1 (LAMC1) (1:8000, 67706-1-Ig; Proteintech Group, Inc., Wuhan, China), collagen type IV alpha-2 chain (COL4A2) (1:5000, 55131-1-AP; Proteintech Group, Inc.), acetyl-CoA acyltransferase 2 (ACAA2) (1:5000, 11111-1-AP; Proteintech Group, Inc.), glutamate-cysteine ligase catalytic subunit (GCLC) (1:6000, 12601-1-AP; Proteintech Group, Inc.), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:100000, 60004-1-Ig; Proteintech Group, Inc.), and α-tubulin (1:6000, PTM-5001; PTM BIO, Hangzhou, China).

Techniques: Biomarker Discovery, Expressing, Standard Deviation

Journal: Rheumatology (Oxford, England)

Article Title: The central helicase domain holds the major conformational epitopes of melanoma differentiation–associated gene 5 autoantibodies

doi: 10.1093/rheumatology/kead397

Figure Lengend Snippet: Determination of melanoma differentiation-associated 5 antibody in patients with DM by cell-based assays and western blot. ( A ) Representative fluorescence microscopy images of CBAs detecting IgG (Hu-IgG) against MDA5 in DM patient (S4 and S22) and healthy control (HC1) serum. A commercial polyclonal MDA5 antibody was used as a positive control. GFP was fused to MDA5 as a control for the expression level. ( B ) WB results of 23 MDA5-Ab-positive DM patient sera in CBAs. ( B ) A heatmap showing MDA5-Ab titres defined by CBAs and the corresponding WB results of the 23 CBAs+ DM patient sera. ( C ) IP assay results of 23 MDA5-Ab-positive DM patient sera in CBAs. GFP: Green Fluorescent Protein; CBAs: cell-based assays

Article Snippet: Patient serum was diluted 1:50, and +99polyclonal anti-MDA5 antibody (Proteintech, #67706–1-Ig) was diluted 1:1000.

Techniques: Western Blot, Fluorescence, Microscopy, Control, Positive Control, Expressing

Journal: Rheumatology (Oxford, England)

Article Title: The central helicase domain holds the major conformational epitopes of melanoma differentiation–associated gene 5 autoantibodies

doi: 10.1093/rheumatology/kead397

Figure Lengend Snippet: Melanoma differentiation-associated protein 5 autoantibody recognizes conformational epitopes. ( A ) Native (left dot) versus denatured (right dot) antigen-based dot blot detecting MDA5-Ab from healthy control, S2, S3, S6, S17 samples were CBAs+/WB+, and S1, S4, S21, S22 samples were CBAs+/WB- DM. Commercial anti-Flag and anti-MDA5 antibodies were used as controls. （ B ） A bar chart showing native (left dot) versus denatured (right dot) antigen-based dot blot detecting MDA5-Ab from healthy control, CBAs+/WB+ samples (S2, S3, S6, S17) and CBAs+/WB- DM samples (S1, S4, S21, S22). * P < 0.05, determined by paired t test. ( C ) Immunoblot (IB) detecting MDA5 by commercial antibody after immunoprecipitation-immunoblot assay of MDA5 using healthy control and CBAs+/WB- DM patient serum. ( D ) MDA5-Ab was determined by antibodies against IgG1. GFP: Green Fluorescent Protein; CBAs: cell-based assays

Article Snippet: Patient serum was diluted 1:50, and +99polyclonal anti-MDA5 antibody (Proteintech, #67706–1-Ig) was diluted 1:1000.

Techniques: Dot Blot, Control, Western Blot, Immunoprecipitation

Journal: Rheumatology (Oxford, England)

Article Title: The central helicase domain holds the major conformational epitopes of melanoma differentiation–associated gene 5 autoantibodies

doi: 10.1093/rheumatology/kead397

Figure Lengend Snippet: Epitope mapping for melanoma differentiation-associated protein 5 using cell-based assays. ( A ) Schematic of constructs coding for full-length MDA5, 2CARD, 3Hel and C-terminal domain domains. ( B ) Representative fluorescence microscopy images of full-length MDA5-, 2CARD-, 3Hel- and C-terminal domain-transfected HEK293 cells immunostained with anti-Flag. ( C ) Representative fluorescence microscopy images of the CBAs detecting IgG against full-length MDA5, 2CARD, 3Hel and C-terminal domain domains in S4 serum. ( D ) Schematic of constructs coding for Hel1, Hel2i and Hel2-pincer domains. ( E ) Representative fluorescence microscopy images of Hel1-, Hel2i- and Hel2-pincer-transfected HEK293 cells immunostained with anti-Flag . ( F ) Representative fluorescence microscopy images of the CBAs detecting IgG against Hel1, Hel2i and Hel2-pincer domains in S4 serum. ( G ) A heatmap showing antibody titres defined by CBAs based on full-length and isolated domains of MDA5. GFP: Green Fluorescent Protein; CBAs: cell-based assays

Article Snippet: Patient serum was diluted 1:50, and +99polyclonal anti-MDA5 antibody (Proteintech, #67706–1-Ig) was diluted 1:1000.

Techniques: Construct, Fluorescence, Microscopy, Transfection, Isolation

Journal: Rheumatology (Oxford, England)

Article Title: The central helicase domain holds the major conformational epitopes of melanoma differentiation–associated gene 5 autoantibodies

doi: 10.1093/rheumatology/kead397

Figure Lengend Snippet: Melanoma differentiation-associated protein 5-autoantibody neutralization by recombinant 3Hel. ( A ) A bar chart showing IFN-β reporter activity in HEK293 cells transfected with poly I: C and either GFP (as a control), full-length MDA5 or 3Hel. Eight samples were detected in the assay; *** P < 0.01 determined by Student’s t -test. ( B ) Immunoblot detecting Flag-tagged recombinant 3Hel purified by immunoprecipitation-immunoblot assay. ( C ) Representative fluorescence microscopy images of the CBAs detecting IgG (Hu-IgG) against MDA5 using serum without (control) and with (immunoadsorbed) preincubation with purified recombinant 3Hel. GFP: Green Fluorescent Protein; CBAs: cell-based assays

Article Snippet: Patient serum was diluted 1:50, and +99polyclonal anti-MDA5 antibody (Proteintech, #67706–1-Ig) was diluted 1:1000.

Techniques: Neutralization, Recombinant, Activity Assay, Transfection, Control, Western Blot, Purification, Immunoprecipitation, Fluorescence, Microscopy

Journal: Rheumatology (Oxford, England)

Article Title: The central helicase domain holds the major conformational epitopes of melanoma differentiation-associated gene 5 autoantibodies.

doi: 10.1093/rheumatology/kead397

Figure Lengend Snippet: Figure 1. Determination of melanoma differentiation-associated 5 antibody in patients with DM by cell-based assays and western blot. (A) Representative fluorescence microscopy images of CBAs detecting IgG (Hu-IgG) against MDA5 in DM patient (S4 and S22) and healthy control (HC1) serum. A commercial polyclonal MDA5 antibody was used as a positive control. GFP was fused to MDA5 as a control for the expression level. (B) WB results of 23 MDA5-Ab-positive DM patient sera in CBAs. (B) A heatmap showing MDA5-Ab titres defined by CBAs and the corresponding WB results of the 23 CBAsþ DM patient sera. (C) IP assay results of 23 MDA5-Ab-positive DM patient sera in CBAs. GFP: Green Fluorescent Protein; CBAs: cell-based assays

Article Snippet: Patient serum was diluted 1:50, and þ99polyclonal anti-MDA5 antibody (Proteintech, #67706–1-Ig) was diluted 1:1000.

Techniques: Western Blot, Microscopy, Control, Positive Control, Expressing

Journal: Rheumatology (Oxford, England)

Article Title: The central helicase domain holds the major conformational epitopes of melanoma differentiation-associated gene 5 autoantibodies.

doi: 10.1093/rheumatology/kead397

Figure Lengend Snippet: Figure 2. Melanoma differentiation-associated protein 5 autoantibody recognizes conformational epitopes. (A) Native (left dot) versus denatured (right dot) antigen-based dot blot detecting MDA5-Ab from healthy control, S2, S3, S6, S17 samples were CBAsþ/WBþ, and S1, S4, S21, S22 samples were CBAsþ/WB- DM. Commercial anti-Flag and anti-MDA5 antibodies were used as controls. (B) A bar chart showing native (left dot) versus denatured (right dot) antigen-based dot blot detecting MDA5-Ab from healthy control, CBAsþ/WBþ samples (S2, S3, S6, S17) and CBAsþ/WB- DM samples (S1, S4, S21, S22). *P < 0.05, determined by paired t test. (C) Immunoblot (IB) detecting MDA5 by commercial antibody after immunoprecipitation-immunoblot assay of MDA5 using healthy control and CBAsþ/WB- DM patient serum. (D) MDA5-Ab was determined by antibodies against IgG1. GFP: Green Fluorescent Protein; CBAs: cell-based assays

Article Snippet: Patient serum was diluted 1:50, and þ99polyclonal anti-MDA5 antibody (Proteintech, #67706–1-Ig) was diluted 1:1000.

Techniques: Dot Blot, Control, Western Blot, Immunoprecipitation

Journal: Rheumatology (Oxford, England)

Article Title: The central helicase domain holds the major conformational epitopes of melanoma differentiation-associated gene 5 autoantibodies.

doi: 10.1093/rheumatology/kead397

Figure Lengend Snippet: Figure 3. Epitope mapping for melanoma differentiation-associated protein 5 using cell-based assays. (A) Schematic of constructs coding for full-length MDA5, 2CARD, 3Hel and C-terminal domain domains. (B) Representative fluorescence microscopy images of full-length MDA5-, 2CARD-, 3Hel- and C- terminal domain-transfected HEK293 cells immunostained with anti-Flag. (C) Representative fluorescence microscopy images of the CBAs detecting IgG against full-length MDA5, 2CARD, 3Hel and C-terminal domain domains in S4 serum. (D) Schematic of constructs coding for Hel1, Hel2i and Hel2-pincer domains. (E) Representative fluorescence microscopy images of Hel1-, Hel2i- and Hel2-pincer-transfected HEK293 cells immunostained with anti-Flag . (F) Representative fluorescence microscopy images of the CBAs detecting IgG against Hel1, Hel2i and Hel2-pincer domains in S4 serum. (G) A heatmap showing antibody titres defined by CBAs based on full-length and isolated domains of MDA5. GFP: Green Fluorescent Protein; CBAs: cell-based assays

Article Snippet: Patient serum was diluted 1:50, and þ99polyclonal anti-MDA5 antibody (Proteintech, #67706–1-Ig) was diluted 1:1000.

Techniques: Construct, Microscopy, Transfection, Isolation

Journal: Rheumatology (Oxford, England)

Article Title: The central helicase domain holds the major conformational epitopes of melanoma differentiation-associated gene 5 autoantibodies.

doi: 10.1093/rheumatology/kead397

Figure Lengend Snippet: Figure 4. Melanoma differentiation-associated protein 5-autoantibody neutralization by recombinant 3Hel. (A) A bar chart showing IFN-b reporter activity in HEK293 cells transfected with poly I: C and either GFP (as a control), full-length MDA5 or 3Hel. Eight samples were detected in the assay; ***P < 0.01 determined by Student’s t-test. (B) Immunoblot detecting Flag-tagged recombinant 3Hel purified by immunoprecipitation-immunoblot assay. (C) Representative fluorescence microscopy images of the CBAs detecting IgG (Hu-IgG) against MDA5 using serum without (control) and with (immunoadsorbed) preincubation with purified recombinant 3Hel. GFP: Green Fluorescent Protein; CBAs: cell-based assays

Article Snippet: Patient serum was diluted 1:50, and þ99polyclonal anti-MDA5 antibody (Proteintech, #67706–1-Ig) was diluted 1:1000.

Techniques: Neutralization, Recombinant, Activity Assay, Transfection, Control, Western Blot, Immunoprecipitation, Microscopy