Journal: Bioconjugate Chemistry

Article Title: Enhancing Reactivity for Bioorthogonal Pretargeting by Unmasking Antibody-Conjugated trans-Cyclooctenes

doi: 10.1021/bc500605g

Figure Lengend Snippet: Figure 5. Translation of dual orthogonal coupling to different antibodies. (A) Anti-TfR and EpCAM antibodies had low TCO reactivity (12−15%) after direct amine-conjugation, but the PEG24− TCO increased reactive TCO density 3- to 4-fold. *MALDI-TOF analysis of the TCO−PEG24-modified anti-EpCAM antibody was limited by difficulties with desorption. (B) Flow cytometry results for TCO- and TCO−PEG24-modified anti-EpCAM and TfR antibodies on A431 cells after reaction with 10 nM tetrazine−QDs for 30 min, showing 5-fold signal enhancements for the PEG24−TCO. (C) Confocal images of A431 cells labeled with (i, iv) anti-EpCAM, (ii, v) anti-TfR, or (iii, vi) nonbinding control antibodies. The antibodies were modified with (i−iii) TCO or (iv−vi) PEG24−TCO at the highest densities and reacted with tetrazine−QD at 10 nM for 30 min. Scale bar: 20 μm. Error bars represent the standard error of at least three independent experiments.

Article Snippet: Monoclonal mouse antibodies specific for human EpCAM (IgG2b, clone 158206) and transferrin receptor (TfR, IgG1, clone 29806) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Conjugation Assay, Flow Cytometry, Labeling, Control

Journal: Nature Aging

Article Title: Iron homeostasis and cell clonality drive cancer-associated intestinal DNA methylation drift in aging

doi: 10.1038/s43587-025-01021-x

Figure Lengend Snippet: a , Schematic representation of a single-crypt DNAm analysis coupled with RNA sequencing. b , Scatter-plot showing the average DNAm level on the Dkk1 , Dkk2 , Sfrp1 and Sfrp2 gene promoters in single crypts isolated from aged mice. The most methylated crypts (the top 10%, high 5mC crypts) and the least-methylated crypts (the bottom 10%, low 5mC crypts) were selected and analyzed by RNA-seq. n = 4 mice were used. c , PCA of single crypt RNA-seq datasets generated in crypts of the top 10% (high 5mC crypts, red dots) and the bottom 10% (low 5mC crypts, blue dots) as in b . To assess the statistical significance of the separation between two groups, a MANOVA test using Pillai’s trace was performed, yielding a highly significant result ( P = 4.235 × 10 −5 ). d , Transferrin receptor ( Tfrc ) and ferroportin ( Slc40a1 ) expression levels in high ( n = 16) and low ( n = 12) 5mC crypts as in b . P value was calculated by a two-tailed Welch’s t -test. Each dot represents a single crypt. n = 4 mice were used. e , Western blot analysis of the protein levels of TfR1 and ferroportin (FPN1) in small intestinal crypts isolated from young and aged mice. Actin was used as a loading control. n = 5 mice per group were analyzed. f , Ferrous (Fe 2+ ) iron levels in small intestinal crypts isolated from young and aged mice. n = 3 mice per group were analyzed. P value was calculated by a two-tailed Welch’s t -test. g , Schematic representation of our hypothesis. h , Quantification of TET hydroxymethylase enzymatic activity in intestinal crypts isolated from young ( n = 5) and aged ( n = 6) mice. P value was calculated by a two-tailed Welch’s t -test. i , Quantification of DNMT enzymatic activity in the intestinal crypts isolated from young ( n = 4) and aged ( n = 5) mice. P value was calculated by a two-tailed Welch’s t -test. j , Bar chart showing DNAm level of the Dkk2 gene promoter analyzed by BS pyrosequencing at the indicated time points upon C35 TET inhibitor treatment of organoids derived from young intestinal crypts. n = 3 mice per group were analyzed. P value was calculated by a two-tailed Welch’s t -tests. k , Bar charts showing DNAm level of the Dkk1 and Dkk2 gene promoters in intestinal crypts isolated from wild-type (WT) and Tet2/3 -dKO mice. n = 3 mice per group were analyzed. P value was calculated by a two-tailed Welch’s t -test. l , Bar chart showing DNAm level of the Dkk2 gene promoter analyzed by BS pyrosequencing in organoids treated with the iron chelator, DFO for 1 and 2 months. n = 3 mice per group were analyzed. P value was calculated by a two-tailed paired t -test. Error bars in the figure bar charts represent the s.d.

Article Snippet: The complementary DNA of murine transferrin receptor ( Tfrc ) was purchased from Origene (plasmid MC221416 ) and cloned into the CSII-EF Venus-P2A-3F-MCS vector using the BamHI and NotI restriction sites.

Techniques: RNA Sequencing, Isolation, Methylation, Generated, Expressing, Two Tailed Test, Western Blot, Control, Activity Assay, Derivative Assay

Journal: Nature Aging

Article Title: Iron homeostasis and cell clonality drive cancer-associated intestinal DNA methylation drift in aging

doi: 10.1038/s43587-025-01021-x

Figure Lengend Snippet: a , The expression level of the Dnmt enzymes in high (red dots; n = 16) and low (blue dots; n = 12) 5mC crypts based on RNAseq as in Fig. . b , The expression level of the Tet enzymes in high (red dots; n = 16) and low (blue dots; n = 12) 5mC crypts based on RNAseq as in Fig. . c , Gene ontology analysis of the differentially expressed genes (DEGs) between the crypts with high (the top 10%) and low (the bottom 10%) DNAm as in Fig. . p-value was calculated by one-sided Fisher’s Exact test. d , Transferrin receptor ( Tfrc ) and ferroportin ( Slc40a1 ) expression levels in the Lgr5 hi cells isolated from young and old mice. n = 3 mice per group were analyzed. p-value is calculated by DESEQ2. e , Quantification of the western blot analysis as in Fig. . n = 5 mice per group were analyzed. p-value was calculated by Welch’s t-test, 2-tails. f , ChIP-qRT-PCR analysis of the ChIP of TET1, TET2 and TET3 on the Dkk2 gene promoter in the intestinal crypts isolated from young and old mice. n = 3 mice per group were analyzed. IgG was used as a control. p-value was calculated by Welch’s t-test, 2-tails. g , ChIP-qRT-PCR analysis of the ChIP of TET1, TET2 and TET3 on the Sfrp1 gene promoter in the intestinal crypts isolated from young and old mice. n = 3 mice per group were analyzed. IgG was used as a control. p-value was calculated by Welch’s t-test, 2-tails. Error bars in the figure bar charts represent the SD.

Article Snippet: The complementary DNA of murine transferrin receptor ( Tfrc ) was purchased from Origene (plasmid MC221416 ) and cloned into the CSII-EF Venus-P2A-3F-MCS vector using the BamHI and NotI restriction sites.

Techniques: Expressing, Isolation, Western Blot, Quantitative RT-PCR, Control

Journal: Nature Aging

Article Title: Iron homeostasis and cell clonality drive cancer-associated intestinal DNA methylation drift in aging

doi: 10.1038/s43587-025-01021-x

Figure Lengend Snippet: a , Western blot analysis of the protein levels of the transferrin receptor (TfR1) in intestinal organoids derived from aged mice and analyzed 40 days after transduction with a 3xFlag-Tfrc- expressing lentivirus. Mock (parental vector) was used as a negative control. Actin was used as a loading control. n = 3 mice per group were analyzed. b , Quantification of TET hydroxymethylase enzymatic activity in intestinal organoids as in a . n = 3 mice per group were analyzed. P value was calculated by a two-tailed Welch’s t -test. c , Bar chart showing the DNAm level of the Dkk2 gene promoter analyzed by BS pyrosequencing in intestinal organoids as in a . Parental organoids, untransduced organoids at day 0. n = 3 mice per group were analyzed. P value was calculated by a two-tailed Welch’s t -test. d , Representative images of the western blot analysis of the protein levels of TfR1 and FPN1 in intestinal organoids cultured in different media as indicated. ENRW (EGF, Noggin, R-spondin1, Wnt3a); ENRC (EGF, Noggin, R-spondin1, CHIR99021). GAPDH was used as a loading control. n = 3 mice per group were analyzed. For details, see the main text. e , Ferrous (Fe 2+) iron levels in intestinal organoids as in d . n = 3 mice per group were analyzed. P value was calculated by one-way analysis of variance (ANOVA). f , Quantification of TET hydroxymethylase enzymatic activity in intestinal organoids as in d . n = 3 mice per group were analyzed. P value was calculated by one-way ANOVA. g , Representative images of the western blot analysis of the protein levels of the TfR1 and FPN1 in intestinal organoids treated with IFNγ (0.2 ng ml −1 ) or with both IFNγ (0.2 ng ml −1 ) and Wnt3a (100 ng ml −1 ) for 5 days as indicated. GAPDH was used as a loading control. n = 3 mice per group were analyzed. For details, see the main text. h , Ferrous (Fe 2+ ) iron levels in intestinal organoids as in g . n = 3 mice per group were analyzed. P value was calculated by one-way ANOVA. i , Quantification of TET hydroxymethylase enzymatic activity in intestinal organoids as in g . n = 3 mice per group were analyzed. P value was calculated by ordinary one-way ANOVA. Error bars in bar charts represent the s.d.

Article Snippet: The complementary DNA of murine transferrin receptor ( Tfrc ) was purchased from Origene (plasmid MC221416 ) and cloned into the CSII-EF Venus-P2A-3F-MCS vector using the BamHI and NotI restriction sites.

Techniques: Western Blot, Derivative Assay, Transduction, Expressing, Plasmid Preparation, Negative Control, Control, Activity Assay, Two Tailed Test, Cell Culture

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: Reagents used in this research.

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques: Comparison, Enzyme-linked Immunosorbent Assay, Diagnostic Assay

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The cross-reaction rates of 9 protein markers with each detection antibody (%).

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques:

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The cross-reaction rates of 9 protein markers with each capture antibody (%).

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques:

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The LLDs and BDLs of liquid protein chip for 9 protein markers (pg/mL).

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques:

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The S -curves of 9 protein markers in the same one detection by using liquid protein chip. ( A ): SF S -curve; ( B ): sTfR S -curve; ( C ): CRP S -curve; ( D ): RBP4 S -curve; ( E ): ApoB S -curve; ( F ): AFP S -curve; ( G ): PA S -curve; ( H ): CEA S -curve; ( I ): D-D S -curve.

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques:

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The regression equations and determination coefficients of 9 protein markers detected by the liquid protein chip.

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques: Marker

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The recovery rates of 9 protein markers detected by liquid protein chip simultaneously ( n = 3).

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques:

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The within-run precisions of 9 protein markers by liquid protein chip ( n = 10).

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques: Chromatin Immunoprecipitation

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The between-run precisions of 9 protein markers by liquid protein chip ( D = 6, n = 3).

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques: Chromatin Immunoprecipitation

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: Correlation analysis between liquid protein chip method and other methods for detecting the 9 protein markers of the same 16 serum samples. ( A ): The correlation analysis of SF; ( B ): The correlation analysis of sTfR; ( C ): The correlation analysis of CRP; ( D ): The correlation analysis of RBP4; ( E ): The correlation analysis of ApoB; ( F ): The correlation analysis of AFP; ( G ): The correlation analysis of PA; ( H ): The correlation analysis of CEA; ( I ): The correlation analysis of D-D.

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques:

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The correlation analysis between the liquid protein chip method and other methods for the detection of 9 protein markers in the same 16 serums ( n = 16).

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The paired comparison between liquid protein chip method and other methods for the detection of 9 protein markers of serums ( n = 16).

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques: Comparison, Enzyme-linked Immunosorbent Assay

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The analytical specificities of different interferences for the liquid protein chip in detecting 9 protein markers (%).

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques:

Journal: Nutrients

Article Title: Construction and Evaluation of a Novel MAP Immunoassay for 9 Nutrition-and-Health-Related Protein Markers Based on Multiplex Liquid Protein Chip Technique

doi: 10.3390/nu15061522

Figure Lengend Snippet: The comparison of the LLDs of 9 protein markers between liquid protein chip and other methods.

Article Snippet: , sTfR detection antiboty , NB100-73092B , Novus Biologicals, Inc. (Centennial, CO, USA).

Techniques: Comparison, Enzyme-linked Immunosorbent Assay