polyclonal antibody against tslp  (R&D Systems)

Journal: NPJ Biofilms and Microbiomes

Article Title: Daily full spectrum light exposure prevents food allergy-like allergic diarrhea by modulating vitamin D 3 and microbiota composition

doi: 10.1038/s41522-021-00213-8

Figure Lengend Snippet: Intestinal levels of IL-25 ( a ), IL-33 ( b ), and thymic stromal lymphopoietin (TSLP) ( c ) were evaluated by quantitative real-time PCR and immunohistochemistry (IHC). Left: values are presented as the mean ± SD of at least eight individuals in each group. *, **, P < 0.05 and 0.01 vs. the control or phototherapy group, respectively (Student’s t test). The pictures at ×400 magnification are representative of at least six individuals in each group. Scale bar: 20 μm.

Article Snippet: After peroxidase blocking, antigen retrieval and blocking, the slides were incubated with rat monoclonal antibody against Mcpt-1 (200× dilution; Thermo Fisher Scientific Inc.), mouse monoclonal antibody against IL-25 (500× dilution; Thermo Fisher Scientific Inc.), mouse monoclonal antibody against IL-33 (200× dilution; Thermo Fisher Scientific Inc.), rabbit polyclonal antibody against TSLP (400× dilution; Thermo Fisher Scientific Inc.), rabbit polyclonal antibody against CYP27A1 (500× dilution; Thermo Fisher Scientific Inc.), or rabbit polyclonal antibody against CYP27B1 (500× dilution; Thermo Fisher Scientific Inc.) overnight at 4 °C followed by incubation with Rat or Mouse/Rabbit Probe HRP (BioTnA, Kaohsiung, Taiwan) for 30 min. After washing, chromogen development was performed by 3,3′-diaminobenzidine staining.

Techniques: Real-time Polymerase Chain Reaction, Immunohistochemistry, Control

Journal: Oncotarget

Article Title: Thymic stromal lymphopoietin in human pancreatic ductal adenocarcinoma: expression and prognostic significance

doi: 10.18632/oncotarget.25997

Figure Lengend Snippet: ( A ) Cytokine mRNA levels in BxPC-3, PT-45 and Capan-2 cells were assessed by real time RT-PCR and normalized to β-actin mRNA levels. Values are means (±SD) normalized gene expression. ( B ) Concentration of TSLP in CM derived from BxPC-3, PT-45, and Capan-2 cells measured by ELISA. The mean level (±SD) of cytokine detected in triplicate in CM samples is indicated.

Article Snippet: Neutralizing polyclonal antibody against TSLP (R&D Systems) or irrelevant control IgG were added to BxPC-3 cell-CM at the concentration used to neutralize TSLP biological activity (40 ng/ml) as reported above.

Techniques: Quantitative RT-PCR, Gene Expression, Concentration Assay, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: Thymic stromal lymphopoietin in human pancreatic ductal adenocarcinoma: expression and prognostic significance

doi: 10.18632/oncotarget.25997

Figure Lengend Snippet: Incidence of OX40L expression on normal HLA-DR + DCs obtained ( A ) in the absence (control), and in the presence of 20% BxPC-3 or Capan-2 cell-CM. Values are means ± SE. P -values obtained by One Way ANOVA followed by Bonferroni t -Test, or ( B ) in the presence of BxPC-3 cell-CM, pre-treated with a neutralizing anti-TSLP polyclonal antibody. Values are means ± SE. P -values obtained by the Paired t -Test.

Article Snippet: Neutralizing polyclonal antibody against TSLP (R&D Systems) or irrelevant control IgG were added to BxPC-3 cell-CM at the concentration used to neutralize TSLP biological activity (40 ng/ml) as reported above.

Techniques: Expressing, Control

Journal: Oncotarget

Article Title: Thymic stromal lymphopoietin in human pancreatic ductal adenocarcinoma: expression and prognostic significance

doi: 10.18632/oncotarget.25997

Figure Lengend Snippet: Basal circulating TSLP levels in PDAC patients ( n = 82) and healthy donors ( n = 41) as determined by ELISA. P -values obtained by the Mann-Whitney Rank Sum Test. Median, 10th, 25th, 75th, and 90th percentiles are presented as vertical boxes with error bars. Dots indicate outliers.

Article Snippet: Neutralizing polyclonal antibody against TSLP (R&D Systems) or irrelevant control IgG were added to BxPC-3 cell-CM at the concentration used to neutralize TSLP biological activity (40 ng/ml) as reported above.

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Oncotarget

Article Title: Thymic stromal lymphopoietin in human pancreatic ductal adenocarcinoma: expression and prognostic significance

doi: 10.18632/oncotarget.25997

Figure Lengend Snippet: Plasma levels of TSLP pre- and post-treatment

Article Snippet: Neutralizing polyclonal antibody against TSLP (R&D Systems) or irrelevant control IgG were added to BxPC-3 cell-CM at the concentration used to neutralize TSLP biological activity (40 ng/ml) as reported above.

Techniques: Clinical Proteomics

Journal: The Journal of investigative dermatology

Article Title: UVB induces HIF-1α-dependent TSLP expression via the JNK and ERK pathways.

doi: 10.1038/jid.2013.203

Figure Lengend Snippet: Figure 1. Upregulation of thymic stromal lymphopoietin (TSLP) expression by UVB irradiation in human keratinocytes. (a) TSLP mRNA levels were upregulated by UVB irradiation in a dose-dependent manner. (b) Elevated TSLP protein expression was detected in a time-dependent manner. (c) UVB irradiation increased TSLP protein secretion in a dose-dependent manner. Secreted TSLP levels in culture supernatant were measured by ELISA. (d) UVB irradiation promoted TSLP expression in the human skin equivalent models (HSEMs). (e) UVB-induced TSLP increased chemokine (c–c motif) ligand 17 (CCL17) secretion by dendritic cells (DCs). CCL17 production in culture supernatant of DCs was determined by ELISA. Bar ¼ 50mm. SB, stratum basale; SC, stratum corneum; SG, stratum granulosum; SP, stratum spinosum. Asterisks indicate statistical significance at *Po0.05, **Po0.01, and ***Po0.001.

Article Snippet: Neutraliz- ing polyclonal antibody against human TSLP was purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Irradiation, Enzyme-linked Immunosorbent Assay

Journal: The Journal of investigative dermatology

Article Title: UVB induces HIF-1α-dependent TSLP expression via the JNK and ERK pathways.

doi: 10.1038/jid.2013.203

Figure Lengend Snippet: Figure 3. Mediation of UVB-induced thymic stromal lymphopoietin (TSLP) upregulation by hypoxia-inducible factor (HIF)-1a in human keratinocytes. (a) TSLP expression was increased by exogenous HIF-1a upregulation at the mRNA and (b) protein levels. To induce the constitutive expression of HIF-1a, pcDNA3–HIF-1a (P402A and P564A) expression vector was transfected into HaCaT keratinocytes, which were then incubated for 24hours. (c) UVB-induced TSLP expression was downregulated by HIF-1a small interfering RNA (siRNA). At about 40% confluence, cells in 60-mm cell culture dishes were transfected with 20nM siRNA and then allowed to stabilize for 24hours before being used in experiments. Scram, scrambled. Asterisks indicate statistical significance at **Po0.01.

Article Snippet: Neutraliz- ing polyclonal antibody against human TSLP was purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Plasmid Preparation, Transfection, Incubation, Small Interfering RNA, Cell Culture

Journal: The Journal of investigative dermatology

Article Title: UVB induces HIF-1α-dependent TSLP expression via the JNK and ERK pathways.

doi: 10.1038/jid.2013.203

Figure Lengend Snippet: Figure 4. Mediation of UVB-induced thymic stromal lymphopoietin (TSLP) upregulation by c-JUN N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) phosphorylation in human keratinocytes. (a) UVB increased mitogen-activated protein kinase (MAPK) phosphorylation in HaCaT keratinocytes. Cells were collected at the different time points after UVB irradiation. JNK, ERK, and p38 were phosphorylated 2 hours after UVB irradiation. (b) Inhibition of MAPKs diminished UVB-induced HIF-1a and TSLP protein expressions. Before UVB irradiation, the cells were pretreated with SP600125, U0126, and SB239063 JNK, ERK, and p38 inhibitors, respectively. Asterisks indicate statistical significance at **Po0.01.

Article Snippet: Neutraliz- ing polyclonal antibody against human TSLP was purchased from R&D Systems (Minneapolis, MN).

Techniques: Phospho-proteomics, Irradiation, Inhibition

Journal: The Journal of investigative dermatology

Article Title: UVB induces HIF-1α-dependent TSLP expression via the JNK and ERK pathways.

doi: 10.1038/jid.2013.203

Figure Lengend Snippet: Figure 5. Regulation of thymic stromal lymphopoietin (TSLP) expression by hypoxia-inducible factor (HIF)-1a through its binding to the TSLP promoter in human keratinocytes. (a) UVB activated the TSLP promoter in HaCaT keratinocytes. (b) HIF-1a activated the TSLP promoter. (c) HIF-1a bound the hypoxia response element (HRE) site of the TSLP promoter. HaCaT keratinocytes were irradiated with UVB and then incubated for 24hours. Chromatin complexes were immunoprecipitated with anti-HIF-1a antibody, and eluted DNA was then amplified by PCR using specific primers. Primers for vascular endothelial growth factor (VEGF) promoter were used as positive control. (d) Arrows show the positions and directions of PCR primers used for detecting the products of the chromatin immunoprecipitation (ChIP) assay. Asterisks indicate statistical significance at *Po0.05.

Article Snippet: Neutraliz- ing polyclonal antibody against human TSLP was purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Binding Assay, Irradiation, Incubation, Immunoprecipitation, Positive Control, Chromatin Immunoprecipitation