rabbit anti pad4  (Proteintech)

Journal: International Journal of Molecular Medicine

Article Title: High-dose X-ray irradiation induces NETosis via the eCIRP/TREM-1 axis in mouse neutrophils

doi: 10.3892/ijmm.2025.5598

Figure Lengend Snippet: Radiation upregulates PAD4 mRNA and protein expression in neutrophils. BMDNs were exposed to 10 Gy radiation. (A) PAD4 mRNA expression levels were assessed 4 h after irradiation via reverse transcription-quantitative PCR. PAD4 protein expression levels were evaluated by western blotting 20 h after irradiation. (B) Representative PAD4 western blot and (C) quantitative bar diagrams are shown. Groups were compared using an unpaired two-tailed Student's t-test. Data are expressed as the mean ± SEM (n=6/group). * P<0.05 and *** P<0.001. Cont, control; Irrad, irradiation; NET, neutrophil extracellular traps; BMDNs, bone marrow-derived neutrophils; PAD4, peptidyl arginine deiminase 4.

Article Snippet: After blocking with 0.1% casein in TBS for 1 h at room temperature, each membrane was incubated overnight at 4°C with primary antibodies of rabbit anti-PAD4 antibody (1:1,000; cat. no. 17373-1-AP; Proteintech Group, Inc.) and mouse anti-β-actin antibody (1:5,000; cat. no. A5441; MilliporeSigma).

Techniques: Expressing, Irradiation, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Two Tailed Test, Control, Derivative Assay

Journal: International Journal of Molecular Medicine

Article Title: High-dose X-ray irradiation induces NETosis via the eCIRP/TREM-1 axis in mouse neutrophils

doi: 10.3892/ijmm.2025.5598

Figure Lengend Snippet: Exposure to ionizing radiation induces NETs via eCIRP/TREM-1 axis. Radiation-induced eCIRP upregulates and activates TREM-1 on neutrophils, resulting in increased expression of PAD4 and NET formation, causing neutropenia. NETs, neutrophil extracellular traps; PAD4, peptidyl arginine deiminase 4; eCIRP, extracellular cold-inducible RNA-binding protein; TREM-1, triggering receptor expressed on myeloid cells-1; MPO, myeloperoxidase; cit, citrulline; CitH3, citrullinated H3. The schema was created in BioRender. Murao, A. (2025) https://BioRender.com/qhaiby0

Article Snippet: After blocking with 0.1% casein in TBS for 1 h at room temperature, each membrane was incubated overnight at 4°C with primary antibodies of rabbit anti-PAD4 antibody (1:1,000; cat. no. 17373-1-AP; Proteintech Group, Inc.) and mouse anti-β-actin antibody (1:5,000; cat. no. A5441; MilliporeSigma).

Techniques: Expressing, RNA Binding Assay

Journal: Pharmaceuticals

Article Title: Effects of the N-Butanol Extract of Pulsatilla Decoction on Neutrophils in a Mouse Model of Ulcerative Colitis

doi: 10.3390/ph17081077

Figure Lengend Snippet: Effects of BEPD on the formation of NETs. ( A – E ) Western blot results of key proteins involved in NET formation, Cit H3, MPO, PAD4, and ELANE. ( ## p < 0.01 vs. control group, * p < 0.05 vs. model group, ** p < 0.01 vs. model group. * p < 0.05 indicates statistically significant).

Article Snippet: The membrane was blocked with 5% non-fat milk powder for 2 h and then the membranes were incubated with rabbit anti-mouse PAD4 (1:1000), rabbit anti-mouse citrullinated histone H3 (Cit H3; 1:1000; catalog number 00057440; Proteintech Group), rabbit anti-mouse MPO (1:5000), and rabbit anti-mouse β-actin (1:10,000; catalog number 380624; Chengdu Zhengneng Biotechnology Co., Ltd.) antibodies overnight at 4 °C.

Techniques: Western Blot, Control

Journal: Pharmaceuticals

Article Title: Effects of the N-Butanol Extract of Pulsatilla Decoction on Neutrophils in a Mouse Model of Ulcerative Colitis

doi: 10.3390/ph17081077

Figure Lengend Snippet: Effects of BEPD on the release of NETs. ( A – E ) IHC staining and semi-quantitative results of colon MPO, MMP-8, MMP-9, PAD4, and ELANE of mice in each group (400× magnification). ( F ) Relative content of MPO, MMP-8, MMP-9, and ELANE in mouse feces. ( # p < 0.05 vs. control group, ## p < 0.01 vs. control group, * p < 0.05 vs. model group, ** p < 0.01 vs. model group. * p < 0.05 indicates statistically significant).

Article Snippet: The membrane was blocked with 5% non-fat milk powder for 2 h and then the membranes were incubated with rabbit anti-mouse PAD4 (1:1000), rabbit anti-mouse citrullinated histone H3 (Cit H3; 1:1000; catalog number 00057440; Proteintech Group), rabbit anti-mouse MPO (1:5000), and rabbit anti-mouse β-actin (1:10,000; catalog number 380624; Chengdu Zhengneng Biotechnology Co., Ltd.) antibodies overnight at 4 °C.

Techniques: Immunohistochemistry, Control

Journal: International journal of molecular sciences

Article Title: Loss of 15-Lipoxygenase in Retinodegenerative RCS Rats.

doi: 10.3390/ijms25042309

Figure Lengend Snippet: Figure 5. ONL PR counts and retinal PAD4 immunofluorescence staining. IVT NAP1051 adminis- tration promotes PR preservation in a dosage-dependent manner (A), which was correlated with PR counts (B). Adjacent histological sections were stained with anti-PAD4 (red) antibodies and DAPI (blue) (C), which showed reduced retinal PAD4 fluorescence intensity (D). Data represented as median ± 95% CI, * p < 0.05, Kruskal–Wallis with Dunn’s correction.

Article Snippet: PAD4 rabbit anti-mouse primary antibody (Proteintech, Sankt Leon-Rot, Germany, Cat. # 17373-1-AP), H3Cit polyclonal rabbit anti-mouse (Abcam, Cambridge, UK, Cat. # ab5103), and MDA polyclonal rabbit anti-mouse (Abcam, Cambridge, UK, Cat. # ab27642) primary antibodies were diluted in blocking buffer and added to each retina and incubated overnight at 4 ◦C.

Techniques: Immunofluorescence, Staining, Preserving, Fluorescence

Journal: International journal of molecular sciences

Article Title: Loss of 15-Lipoxygenase in Retinodegenerative RCS Rats.

doi: 10.3390/ijms25042309

Figure Lengend Snippet: Figure 9. Effects on rd10 mice treated with NAP1051. rd10 mice treated with NAP1051 from p15 to p26 showed ONL preservation on OCT (arrows) and H&E on p26 (A). NAP1051 was able to preserve ONL thickness (B) and PR counts (C). OCT-FAF showed NAP1051 reduced hyperpigmentation on p26 (D,E). Correlations between the ONL PR counts and FAF hyperpigmentation were evaluated and demonstrated a negative linear correlation (R2 = 0.9) where fewer hyperpigmentation spots were correlated to increased ONL PR nuclei (F). Retinal PAD4 immunofluorescence staining was reduced for animals treated with 37.5 mg/kg/BID (G) corresponding with reduced retinal H3Cit, but not to a statistically significant level (H). The oxidative stress marker malondialdehyde (MDA) was statisti- cally significantly reduced at the 37.5 mg/kg/BID dosage (I). Data represented as median ± 95% CI, * p < 0.05, ** p < 0.01, Kruskal–Wallis test with Dunn’s correction. The number of PRs in the ONL and hyperpigmentation spots seen in FAF were enumerated using ImageJ/FIJI (ver 2.3.0).

Article Snippet: PAD4 rabbit anti-mouse primary antibody (Proteintech, Sankt Leon-Rot, Germany, Cat. # 17373-1-AP), H3Cit polyclonal rabbit anti-mouse (Abcam, Cambridge, UK, Cat. # ab5103), and MDA polyclonal rabbit anti-mouse (Abcam, Cambridge, UK, Cat. # ab27642) primary antibodies were diluted in blocking buffer and added to each retina and incubated overnight at 4 ◦C.

Techniques: Preserving, Immunofluorescence, Staining, Marker