Journal: Frontiers in Immunology

Article Title: Characterization of circulating extracellular traps and immune responses to citrullinated LL37 in psoriasis

doi: 10.3389/fimmu.2023.1247592

Figure Lengend Snippet: Antibodies used for detection of LL37, citLL37 and IgG in circulating NETs.

Article Snippet: Rabbit anti-human serum albumin , 1.33 μg/ml (cat: 109-4133; Rockland) , HRP-goat anti-rabbit IgG , 0.067 μg/ml (cat: P0448, Dako).

Techniques: Concentration Assay

Journal:

Article Title: Survival of Aspergillus fumigatus in Serum Involves Removal of Iron from Transferrin: the Role of Siderophores

doi: 10.1128/IAI.72.3.1402-1408.2004

Figure Lengend Snippet: Degradation of transferrin by A. fumigatus in liquid cultures. A. fumigatus was incubated in MEM containing 10% human serum (A) or 2.5 μM holotransferrin (B). Supernatants were withdrawn from the cultures after the number of hours indicated above the lanes, and the presence of transferrin was determined by Western blotting following sodium dodecyl sulfate-PAGE. Controls (lanes C) were uninoculated samples incubated for 286 h. The band underneath transferrin in panel A is another protein that cross-reacted with the polyclonal antitransferrin antibody.

Article Snippet: Gels were silver stained or transferred to polyvinylidene difluoride membranes (Bio-Rad), blocked with 5% bovine serum albumin, probed with a rabbit immunoglobulin G fraction of anti-human transferrin (1:1,000 dilution; Rockland Inc., Gilbertsville, Pa.), and treated with goat anti-rabbit horseradish peroxidase.

Techniques: Incubation, Western Blot

Journal:

Article Title: Survival of Aspergillus fumigatus in Serum Involves Removal of Iron from Transferrin: the Role of Siderophores

doi: 10.1128/IAI.72.3.1402-1408.2004

Figure Lengend Snippet: Iron removal from transferrin by A. fumigatus. A. fumigatus was cultured in MEM containing 2.5 μM purified human holotransferrin (A) or 10% human serum (B). Culture media were withdrawn, and the iron saturation of transferrin was analyzed by urea-PAGE. Transferrin was visualized by Western blotting. The numbers above the lanes represent the hours of incubation with A. fumigatus. Fe2-Tf, holotransferrin; Apo-Tf, apotransferrin.

Article Snippet: Gels were silver stained or transferred to polyvinylidene difluoride membranes (Bio-Rad), blocked with 5% bovine serum albumin, probed with a rabbit immunoglobulin G fraction of anti-human transferrin (1:1,000 dilution; Rockland Inc., Gilbertsville, Pa.), and treated with goat anti-rabbit horseradish peroxidase.

Techniques: Cell Culture, Purification, Western Blot, Incubation

Journal:

Article Title: Survival of Aspergillus fumigatus in Serum Involves Removal of Iron from Transferrin: the Role of Siderophores

doi: 10.1128/IAI.72.3.1402-1408.2004

Figure Lengend Snippet: A. fumigatus can transport iron from transferrin across a dialysis membrane. A. fumigatus was inoculated into MEM in which a dialysis bag containing holotransferrin (25 μM) was suspended. A. fumigatus was incubated for 48 h, and then transferrin was withdrawn from the dialysis bag and analyzed by urea-PAGE (lane +). An uninoculated control flask containing MEM plus transferrin in a dialysis bag also was examined (lane −). Pure holotransferrin (Fe2-Tf) and apotransferrin (Apo-Tf) standards also were run.

Article Snippet: Gels were silver stained or transferred to polyvinylidene difluoride membranes (Bio-Rad), blocked with 5% bovine serum albumin, probed with a rabbit immunoglobulin G fraction of anti-human transferrin (1:1,000 dilution; Rockland Inc., Gilbertsville, Pa.), and treated with goat anti-rabbit horseradish peroxidase.

Techniques: Incubation

Journal:

Article Title: Survival of Aspergillus fumigatus in Serum Involves Removal of Iron from Transferrin: the Role of Siderophores

doi: 10.1128/IAI.72.3.1402-1408.2004

Figure Lengend Snippet: Iron saturation of transferrin following incubation with A. fumigatus siderophores. Purified desferrisiderophores were incubated with holotransferrin (25 μM) at 37°C for 16 h. Desferriferrichrome, desferriferricrocin, and desferritriacetylfusarinine C were serially diluted to final concentrations of 5 μM (lanes 1), 50 μM (lanes 2), 500 μM (lanes 3), and 5 mM (lanes 4). Controls containing holotransferrin alone also were run (lanes 0). holo-Tf, holotransferrin; Fe-Tf, monoferric transferrin; apo-Tf, apotransferrin.

Article Snippet: Gels were silver stained or transferred to polyvinylidene difluoride membranes (Bio-Rad), blocked with 5% bovine serum albumin, probed with a rabbit immunoglobulin G fraction of anti-human transferrin (1:1,000 dilution; Rockland Inc., Gilbertsville, Pa.), and treated with goat anti-rabbit horseradish peroxidase.

Techniques: Incubation, Purification

Journal:

Article Title: Base Excision Repair Is Limited by Different Proteins in Male Germ Cell Nuclear Extracts Prepared from Young and Old Mice

doi: 10.1128/MCB.22.7.2410-2418.2002

Figure Lengend Snippet: Western blot analysis of BER proteins in nuclear extracts prepared from enriched populations of primitive type A spermatogonia (PA), type A spermatogonia (A), type B spermatogonia (B), pachytene spermatocytes (P), and round spermatids (R). Bands corresponding to DNA ligase I (130 kDa), DNA ligase III (93 kDa), Xrcc-1 (69 kDa), β-pol (39 kDa), and Ape/Ref-1 (37 kDa) proteins were visualized. Triplicate assays for each of three independent nuclear extract preparations were performed. Some variation between replicates was noted. A summary of all data is shown in Table ​Table1.1. Molecular mass protein standards and purified DNA ligases I and III, β-pol, and Ape/Ref-1 (lane STD) are shown for comparison.

Article Snippet: Rabbit anti-human APE/REF-1 (hAPE/REF-1; Novus Biologicals, Littleton, Colo.) and rabbit polyclonal anti-β-pol (S. Wilson, NIEHS, Research Triangle Park, N.C.) were used to detect Ape/Ref-1 and β-pol, respectively.

Techniques: Western Blot, Purification, Comparison

Journal:

Article Title: Base Excision Repair Is Limited by Different Proteins in Male Germ Cell Nuclear Extracts Prepared from Young and Old Mice

doi: 10.1128/MCB.22.7.2410-2418.2002

Figure Lengend Snippet: Western blot analysis of BER proteins in MGC nuclear extracts prepared from 3-month-old, 16-month-old, and 28-month-old mice. Bands corresponding to DNA ligase I (130 kDa), DNA ligase III (93 kDa), Xrcc-1 (69 kDa), β-pol (39 kDa), and Ape/Ref-1 (37 kDa) proteins were visualized. Triplicate assays were performed for each of three independent nuclear extract preparations. Some variation in signal intensities was observed among Western blots. A summary of all data is shown in Table ​Table2.2. Molecular mass protein standards and purified DNA ligases I and III, β-pol, and APE/REF-1 (lane STD) are shown for comparison.

Article Snippet: Rabbit anti-human APE/REF-1 (hAPE/REF-1; Novus Biologicals, Littleton, Colo.) and rabbit polyclonal anti-β-pol (S. Wilson, NIEHS, Research Triangle Park, N.C.) were used to detect Ape/Ref-1 and β-pol, respectively.

Techniques: Western Blot, Purification, Comparison

Journal:

Article Title: Base Excision Repair Is Limited by Different Proteins in Male Germ Cell Nuclear Extracts Prepared from Young and Old Mice

doi: 10.1128/MCB.22.7.2410-2418.2002

Figure Lengend Snippet: UDG-BER activities for MGC nuclear extracts prepared from 3-month-old B6D2F1 (open), 28-month-old B6D2F1 (solid), and 3-month-old Apex heterozygous knockout mice (striped). Results are presented as means ± SEM of three replicate assays for each of three independent nuclear extract preparations. ∗, significantly (P < 0.05) different from addition of 0 ng of protein within a specific group. †, amount of purified APE/REF-1 required to restore activity to that of 3-month-old mice with 0 ng of protein added.

Article Snippet: Rabbit anti-human APE/REF-1 (hAPE/REF-1; Novus Biologicals, Littleton, Colo.) and rabbit polyclonal anti-β-pol (S. Wilson, NIEHS, Research Triangle Park, N.C.) were used to detect Ape/Ref-1 and β-pol, respectively.

Techniques: Knock-Out, Purification, Activity Assay

Journal: Cancer biology & therapy

Article Title: The combination of 5-fluorouracil plus p53 pathway restoration is associated with depletion of p53-deficient or mutant p53-expressing putative colon cancer stem cells.

doi: 10.4161/cbt.8.22.10446

Figure Lengend Snippet: Figure 1. Upregulation of p53-dependent transcriptional activity or endogenous pro-apoptotic TRAIL receptor DR5 expression following p53 pathway restoration. In the upper panels, a whole population of Phoenix cells expressing a p53-responsive EGFP reporter show increased EGFP reporter expression following infection by a p73-expressing Adenovirus (upper right vs. upper left). In the lower panels, mutant p53-expressing human colon cancer DLD1 cells were treated with 15 μg/ml of the mutant p53 conformation modifying drug CP-31398 for 12 hours followed by fluorescence microscopy for p53 target TRAIL receptor DR5 expression by immunofluorescence with a DAPI nuclear counterstain. The lower left panel shows DR5 expression in untreated DLD1 cells and the lower right panel shows DR5 expression in CP-31398 treated DLD1 cells.

Article Snippet: A rabbit anti-human DR5 IgG antibody (Imgenex) was added at a 1:100 dilution and this was incubated overnight with gentle shaking at 4°C.

Techniques: Activity Assay, Expressing, Infection, Mutagenesis, Fluorescence, Microscopy, Immunofluorescence

Journal: iScience

Article Title: DNA methylation of the IL1R2 gene is associated with porcine placental development and birth weight

doi: 10.1016/j.isci.2026.115055

Figure Lengend Snippet: RT-qPCR analysis of DNMTs and IL1R2 expression, along with IHC staining (A) RT-qPCR was used to quantitatively analyze the expression profiles of DNMTs and IL1R2 in HBW and LBW placentas ( DNMT3A : p = 0.0264; DNMT3L : p = 0.0016; DNMT1 : p = 0.0069; IL1R2 : p = 0.0156). (B and D) IHC was performed to analyze the protein expression intensity of DNMT3A in HBW and LBW placentas (DNMT3A: p = 0.0059). Scale bars, 50 μm. (C and E) IHC was performed to analyze the protein expression intensity of IL1R2 in HBW and LBW placentas (IL1R2: p = 0.0007). Scale bars, 50 μm. Data are represented as mean ± SEM. Statistical significance was assessed by paired Student’s t test, with n = 6 (3 samples in the LBW group and 3 samples in the HBW group).

Article Snippet: Rabbit anti-Homo sapiens (Human) IL1R2 Polyclonal antibody , Cusabio Biotech , Cat# CSB-PA011622ESR1HU; RRID: AB_3719906.

Techniques: Quantitative RT-PCR, Expressing, Immunohistochemistry

Journal: iScience

Article Title: DNA methylation of the IL1R2 gene is associated with porcine placental development and birth weight

doi: 10.1016/j.isci.2026.115055

Figure Lengend Snippet: MethPrimer-based prediction of CpG islands in the IL1R2 promoter region and subsequent analysis of the methylation status in placental tissues (A) Bioinformatics analysis predicted CpG islands in the promoter region of IL1R2 . (B) MS-PCR measured IL1R2 methylation levels in HBW and LBW placentas ( p = 0.0062). Data are represented as mean ± SEM. Statistical significance was assessed by unpaired Student’s t test, with n = 6 (3 samples in the LBW group and 3 samples in the HBW group).

Article Snippet: Rabbit anti-Homo sapiens (Human) IL1R2 Polyclonal antibody , Cusabio Biotech , Cat# CSB-PA011622ESR1HU; RRID: AB_3719906.

Techniques: Methylation

Journal: iScience

Article Title: DNA methylation of the IL1R2 gene is associated with porcine placental development and birth weight

doi: 10.1016/j.isci.2026.115055

Figure Lengend Snippet: Effects of 5-Aza treatment on IL1R2 gene expression and methylation levels in PTr2 cells (A) The mRNA expression of IL1R2 in 5-Aza-treated PTr2 cells was measured using RT-qPCR (20 μM: p = 0.0199). “ns” denotes non-significant results ( p > 0.05). (B) MS-PCR measured the methylation levels of the IL1R2 promoter in 5-Aza-treated PTr2 Cells ( p = 0.0005). (C) BS-PCR measured the methylation levels of the IL1R2 promoter in 5-Aza-treated PTr2 cells ( p = 0.0011). Data are represented as mean ± SEM. Statistical significance was assessed by unpaired Student’s t test, n = 3.

Article Snippet: Rabbit anti-Homo sapiens (Human) IL1R2 Polyclonal antibody , Cusabio Biotech , Cat# CSB-PA011622ESR1HU; RRID: AB_3719906.

Techniques: Gene Expression, Methylation, Expressing, Quantitative RT-PCR

Journal: iScience

Article Title: DNA methylation of the IL1R2 gene is associated with porcine placental development and birth weight

doi: 10.1016/j.isci.2026.115055

Figure Lengend Snippet: IL1R2 knockdown promotes PTr2 Cell proliferation and migration (A) RT-qPCR analysis of IL1R2 knockdown efficiency and its effect on proliferation and apoptosis markers in PTr2 cells ( IL1R2 : p = 0.0002; Ki67 : p = 0.0043; BAX : p = 0.0155; CASP3 : p = 0.0088; CASP9 : p = 0.0331). “ns” denotes non-significant results ( p > 0.05). (B) Cell proliferation assessed using CCK-8 assay after IL1R2 knockdown (48 h: p = 0.0224; 72 h: p = 0.0035). (C and D) Cell proliferation evaluated using EdU assay ( p = 0.0083). Scale bars, 100 μm. (E and F) Cell migration was analyzed by scratch assay ( p = 0.0462). Scale bars, 500 μm. Data are represented as mean ± SEM. Statistical significance was assessed by unpaired Student’s t test, n = 3.

Article Snippet: Rabbit anti-Homo sapiens (Human) IL1R2 Polyclonal antibody , Cusabio Biotech , Cat# CSB-PA011622ESR1HU; RRID: AB_3719906.

Techniques: Knockdown, Migration, Quantitative RT-PCR, CCK-8 Assay, EdU Assay, Wound Healing Assay

Journal: iScience

Article Title: DNA methylation of the IL1R2 gene is associated with porcine placental development and birth weight

doi: 10.1016/j.isci.2026.115055

Figure Lengend Snippet: IL1R2 overexpression inhibits PTr2 Cell proliferation and migration (A) RT-qPCR analysis of IL1R2 overexpression efficiency and its effect on proliferation and apoptosis markers in PTr2 cells ( IL1R2 : p = 0.0003; PCNA : p = 0.0256; BAX : p = 0.0497). “ns” denotes non-significant results ( p > 0.05). (B) Cell proliferation was assessed using CCK-8 assay following IL1R2 overexpression (48 h: p = 0.0027; 72 h: p = 0.0011). (C and D) Cell proliferation evaluated using EdU assay ( p = 0.0452). Scale bars, 100 μm. (E and F) Cell migration was analyzed by scratch assay ( p = 0.0078). Scale bars, 500 μm. Data are represented as mean ± SEM. Statistical significance was assessed by unpaired Student’s t test, n = 3.

Article Snippet: Rabbit anti-Homo sapiens (Human) IL1R2 Polyclonal antibody , Cusabio Biotech , Cat# CSB-PA011622ESR1HU; RRID: AB_3719906.

Techniques: Over Expression, Migration, Quantitative RT-PCR, CCK-8 Assay, EdU Assay, Wound Healing Assay

Journal: iScience

Article Title: DNA methylation of the IL1R2 gene is associated with porcine placental development and birth weight

doi: 10.1016/j.isci.2026.115055

Figure Lengend Snippet: IL1R2 positively regulates TNF- α expression in PTr2 cells (A) IL1R2 knockdown suppresses TNF-α mRNA levels ( p = 0.0442). (B) IL1R2 overexpression elevates TNF-α mRNA levels ( p = 0.0057). (C - E) IL1R2 knockdown reduces TNF- α protein abundance, with confirmation of knockdown efficiency (IL1R2: p = 0.0012; TNF-α: p = 0.0394). (F - H) IL1R2 overexpression increases TNF- α protein levels, with confirmation of knockdown efficiency (IL1R2: p = 0.0011; TNF-α: p = 0.0014). Data are represented as mean ± SEM. Statistical significance was assessed by unpaired Student’s t test, n = 3.

Article Snippet: Rabbit anti-Homo sapiens (Human) IL1R2 Polyclonal antibody , Cusabio Biotech , Cat# CSB-PA011622ESR1HU; RRID: AB_3719906.

Techniques: Expressing, Knockdown, Over Expression, Quantitative Proteomics