polyclonal rabbit anti il 1ra antibody  (Proteintech)

Journal: BMC Cancer

Article Title: Bladder cancer-derived interleukin-1 converts the vascular endothelium into a pro-inflammatory and pro-coagulatory surface

doi: 10.1186/s12885-020-07548-z

Figure Lengend Snippet: Pro-inflammatory activation of endothelial cells after stimulation with tumour cell SN. a After incubation with T24 cell SN, CXCL-1, IL-6, IL-8, GM-CSF and PAI-1 were significantly released from HUVECs (HUVECs + T24 cell SN). The SN of UROtsa was not able to stimulate HUVECs (HUVECs + UROtsa SN). HUVECs: baseline cytokine levels produced by HUVECs; T24 and UROTsa cell SN: baseline levels of cytokine produced by the tumour cells; ( n = 8–12) ** P ≤ 0.01. b Incubation of HUVECs for 6 h with T24 cell SN promoted the trans-localization of NF-kB (red) into the nucleus (blue) of the endothelial cells. Treatment of HUVECs with the SN of RT112 and UROtsa cells had no effect. RT4 cell SN induced a weak trans-localization of NF-kB. Addition of IL-1ra blocked NF-kB translocation upon treatment with T24 cell SN. The bar diagram shows the fluorescence intensity of nuclear NF-kB. Scale bar corresponds to 50 μm; ( n = 10 images) ** P ≤ 0.01. c IL-1ra inhibited the T24 cell SN induced release of the indicated pro-inflammatory cytokines from HUVECs. Cytokine levels were shown in pg/ml; ( n = 3) * P ≤ 0.05

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-human IL-1ra antibody (Sigma-Aldrich, St. Louis, MO, USA) 1:200 in DAKO Real antibody diluent (DAKO, Glostrup, Denmark).

Techniques: Activation Assay, Incubation, Produced, Translocation Assay, Fluorescence

Journal: BMC Cancer

Article Title: Bladder cancer-derived interleukin-1 converts the vascular endothelium into a pro-inflammatory and pro-coagulatory surface

doi: 10.1186/s12885-020-07548-z

Figure Lengend Snippet: Impact of UBC cell SN on the integrity of the endothelial cell layer. a HUVECs grown on gelatine coated ECIS electrodes were treated with starvation medium (control) or UBC cell SN. The impedance was measured continuously up to 12 h after treatment. T24 cell SN led to marked breakdown of the endothelial impedance. RT4 cell SN had a lower impact whereas RT112 cell SN, UROtsa cell SN or starvation medium (control) did not cause significant alterations after 12 h; n = 5; ** P ≤ 0.01. b HUVEC cells were incubated for 12 h with T24 cell SN (T24) or starvation medium (control) and then analysed by immunofluorescence staining of β-Actin (red) and CD31/PECAM-1 (green). In comparison to the control (left), treatment with T24 cell SN (right) induced the disintegration of adherent junctions (CD31/PECAM-1) indicating a weakening of the endothelial barrier. c Inhibition of cytokine signalling partially conserved endothelial barrier function. Antibodies against IL-6, CXCL-1 slightly mitigated impedance breakdown. In contrast, IL-1ra strongly attenuated tumour mediated endothelial dysfunction, whereas IL-8 neutralization had no impact; n = 4; ** P ≤ 0.01

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-human IL-1ra antibody (Sigma-Aldrich, St. Louis, MO, USA) 1:200 in DAKO Real antibody diluent (DAKO, Glostrup, Denmark).

Techniques: Incubation, Immunofluorescence, Staining, Inhibition, Neutralization

Journal: BMC Cancer

Article Title: Bladder cancer-derived interleukin-1 converts the vascular endothelium into a pro-inflammatory and pro-coagulatory surface

doi: 10.1186/s12885-020-07548-z

Figure Lengend Snippet: Impact of IL-1 on bladder cancer progression. a IL-1ra staining intensity in a cohort of 105 patients with UBC measured semiquantitatively by IHC. Controls (benign urothelium) showed a higher IL1-ra expression than in patients with cancer. No significant difference was found between low grade and high grade tumours. The staining intensity score was defined as follows: no staining = 1, low intensity = 2, moderate intensity = 3 and high intensity = 4. * P ≤ 0.05. n.s. = not significant. Representative IHC images are shown in b - c . Scale bar corresponds to 500 μm. b IHC very low staining intensity (score = 1). c Intermediate staining intensity (score = 3). d High staining intensity (score = 4). e High IL-1β mRNA levels were linked to adverse clinicopathological features high grade disease, muscle invasiveness and tumour progression. MIBC: muscle invasive bladder cancer, NMIBC: non-muscle invasive bladder cancer; * P ≤ 0.05

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-human IL-1ra antibody (Sigma-Aldrich, St. Louis, MO, USA) 1:200 in DAKO Real antibody diluent (DAKO, Glostrup, Denmark).

Techniques: Staining, Expressing

Journal: Molecular Metabolism

Article Title: Amyloid formation disrupts the balance between interleukin-1β and interleukin-1 receptor antagonist in human islets

doi: 10.1016/j.molmet.2017.05.016

Figure Lengend Snippet: IL-1Ra levels are increased in oligomer-positive islet areas and reduced in thioflavin S-positive islet areas. (A) Paraffin embedded sections from 7-day cultured (11.1 mmol/L glucose) non-transduced or transduced with Ad-prohIAPP-siRNA (or Ad-control-siRNA) islets were immunolabeled for insulin (red) and IL-1Ra (green). The squares (dashed white lines) denote enlarged regions shown as inserts. (B) Human islets were immunolabeled for IL-1Ra (green) and A11 (red; top panel) or insulin (red), IL-1Ra (green) and thioflavin S (Thio S; blue; bottom panel). (C) Paraffin-embedded sections from (left to right): ob/ob mouse and human adipose tissue immunolabeled for IL-1Ra (green; positive control); human islets incubated with secondary antibody alone (negative control) and anakinra-treated human islets immunolabeled for insulin (red) and IL-1Ra (green). Scale bar = 50 μm; inserts: ×3 (A11: ×4). Micrographs are representative of four independent studies (4 human islet preparations).

Article Snippet: IL-1Ra immuno-labeling was performed using a polyclonal rabbit anti-IL-1Ra antibody (ThermoFisher; PA5-21776).

Techniques: Cell Culture, Transduction, Control, Immunolabeling, Positive Control, Incubation, Negative Control

Journal: Molecular Metabolism

Article Title: Amyloid formation disrupts the balance between interleukin-1β and interleukin-1 receptor antagonist in human islets

doi: 10.1016/j.molmet.2017.05.016

Figure Lengend Snippet: IL-1Ra immunoreactivity is detectable in human islet α-cells. (A) Paraffin-embedded sections from human islets transduced with Ad-prohIAPP-siRNA (or Ad-control-siRNA) and cultured (11.1 mmol/L glucose) for 7 days were immunolabeled for glucagon (red) and IL-1Ra (green). (B) Triple immunostaining of pre-culture human islets for insulin (blue), glucagon (red) and IL-1Ra (green). The squares (dashed white lines) denote regions enlarged and depicted as inserts at the bottom right of the corresponding images. Amyloid formation during islet culture is shown as an insert in merged micrographs (top right, insulin; red and thioflavin S; blue). Scale bar = 50 μm; inserts: ×3. Micrographs are representative of four independent studies (4 human islet preparations).

Article Snippet: IL-1Ra immuno-labeling was performed using a polyclonal rabbit anti-IL-1Ra antibody (ThermoFisher; PA5-21776).

Techniques: Transduction, Control, Cell Culture, Immunolabeling, Triple Immunostaining

Journal: Molecular Metabolism

Article Title: Amyloid formation disrupts the balance between interleukin-1β and interleukin-1 receptor antagonist in human islets

doi: 10.1016/j.molmet.2017.05.016

Figure Lengend Snippet: IL-1Ra release from Ad-prohIAPP-siRNA transduced and non-transduced human islets cultured with or without neutralizing IL-1β antibody. (A, B) IL-1Ra release from human islets transduced with Ad-prohIAPP-siRNA or Ad-control-siRNA following 3 or 7 days culture (11.1 mmol/L glucose) in the presence (+nIL1β) or absence (-nIL1β) of neutralizing IL-1β antibody. (C) Comparison between islet IL-1Ra release following 3 and 7 days culture in each condition. Data are expressed as mean ± SEM of four independent studies (4 human islet preparations) performed in duplicate. *vs corresponding day 3 group ( p < 0.05, two-way ANOVA).

Article Snippet: IL-1Ra immuno-labeling was performed using a polyclonal rabbit anti-IL-1Ra antibody (ThermoFisher; PA5-21776).

Techniques: Cell Culture, Transduction, Control, Comparison

Journal: Molecular Metabolism

Article Title: Amyloid formation disrupts the balance between interleukin-1β and interleukin-1 receptor antagonist in human islets

doi: 10.1016/j.molmet.2017.05.016

Figure Lengend Snippet: A proposed mechanism for islet amyloid-induced islet inflammation and β-cell apoptosis. Amyloid formation promotes islet IL-1β production while reducing islet IL-1Ra levels resulting in increased IL-1β/IL-1Ra ratio thereby contributing to islet inflammation, β-cell dysfunction, and apoptosis. Blocking IL-1β signaling or preserving islet IL-1Ra levels to restore IL-1β/IL-1Ra balance may provide effective potential strategies to protect islet β-cells from amyloid toxicity in pathologic conditions associated with islet amyloid formation.

Article Snippet: IL-1Ra immuno-labeling was performed using a polyclonal rabbit anti-IL-1Ra antibody (ThermoFisher; PA5-21776).

Techniques: Blocking Assay, Preserving