Human/Mouse NLRP3/NALP3 Antibody

Detection of Mouse NLRP3/NALP3 by Western Blot. Western blot shows lysates of J774A.1 mouse reticulum cell sarcoma macrophage cell line and RAW 264.7 mouse monocyte/macrophage cell line. PVDF membrane was probed with 2 µg/mL of Rat Anti-Human/Mouse NLRP3/NALP3 Monoclonal Antibody (Catalog # MAB7578) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for NLRP3/NALP3 at approximately 117 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.Catalog # AF6789is recommended for use with human lysates.

NLRP3/NALP3 in RAW 264.7 Mouse Cell Line. NLRP3/NALP3 was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Rat Anti-Human/Mouse NLRP3/NALP3 Monoclonal Antibody (Catalog # MAB7578) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

NLRP3/NALP3 in Human Tonsil Tissue. NLRP3/NALP3 was detected in immersion fixed paraffin-embedded sections of human tonsil tissue using Rat Anti-Human/Mouse NLRP3/NALP3 Monoclonal Antibody (Catalog # MAB7578) at 5 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Rat IgG VisUCyte™ HRP Polymer Antibody (brown; Catalog # VC005) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of NLRP3/NALP3 in RAW 264.7 Mouse Cell Line by Flow Cytometry. RAW 264.7 mouse monocyte/macrophage cell line was stained with Rat Anti-Human/Mouse NLRP3/ NALP3 Monoclonal Antibody (Catalog # MAB7578, filled histo-gram) or isotype control antibody (Catalog # MAB006, open histo-gram), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Detection of NLRP3/NALP3 in Human Monocytes by Flow Cytometry. Human peripheral blood monocytes was stained with Rat Anti-Human/Mouse NLRP3/NALP3 Monoclonal Antibody (Catalog # MAB7578, filled histogram) or isotype control antibody (Catalog # MAB006, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Detection of Mouse NLRP3/NALP3 by Western Blot Sodium butyrate (NaB) and hydroxybutyric acid (HBA) eyedrops improve clinical parameters and decrease inflammation in animals subjected to alkali burns (AB). (A) Digital images of corneas stained with 0.1% sodium fluorescein demonstrating intact corneal epithelium after topical NaB, HBA or balanced salt solution (BSS) treatment for 5 days quater in die (QID) in untreated mice (n = 5 animals/group). Scale bar: 1000 µm; (B) Representative color digital images used to score corneal opacity (top row) and representative fluorescein stained corneas used to create wound closure rate (bottom row) at 5 days (n = 15 animals/group) after alkali burn. Scale bar: 1000 µm; (C) Corneal opacity in corneas subjected to alkali burn and topically treated with either NaB, HBA, or BSS and compared with dexamethasone (Dex) (n = 15 animals/group); (D) Wound closure rate in corneas subjected to alkali burn and topically treated with either NaB, HBA, or BSS and compared with Dex (n = 15 animals/group); (E) Mean ± SEM of results of gene expression analysis of NLRP3, Caspase-1, and IL-1 beta in whole cornea of animals subjected to alkali burn for 2 or 5 days and topically treated with either NaB, HBA, or BSS and compared with Dex (n = 5 animals/group). * p < 0.05, *** p < 0.001, **** p < 0.0001. (F) Representative digital images of western blot of NLRP3 and beta -actin in cornea epithelium of animals subjected to alkali burn for 2 days and topically treated with either NaB, HBA, or BSS (n = 12 animals/3 samples/group). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28273882), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse NLRP3/NALP3 by Western Blot Upregulation of nucleotide-binding oligomerization domain-containing protein (NOD)-like receptor family pyrin domain containing 3 (NLRP3) expression by corneal epithelium in animals subjected to alkali burn (AB) at 2 days (2 d) and 5 days (5 d) post-injury. (A) Gene expression analysis of NLRP3 messenger mRNA (mRNA) transcript in whole cornea of animals subjected to alkali burn (n = 8 animals/group). Graphs show means ± standard error of the mean (SEM). **** p < 0.0001; (B) Representative merged digital images of laser scanning confocal microscopy of corneas cryosections immunostained for NLRP3 (green) with propidium iodide nuclei counterstaining (DNA in red) in corneas subjected to alkali burn (images are representative of n = 6 animals/group). Scale bar: 50 µm; (C) Representative digital images of western blot of NLRP3 and beta -actin in cornea epithelium of animals subjected to alkali burn for 2 and 5 days. Each lane is a different sample (n = 12 animals/3 samples/group). MW: Molecular weight; UT: untreated control; 2 d and 5 d refer to mice subjected to alkali burn and euthanized 2 or 5 days post-injury, respectively (1 and 2 indicate different samples). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28273882), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse NLRP3/NALP3 by Immunocytochemistry/Immunofluorescence Upregulation of nucleotide-binding oligomerization domain-containing protein (NOD)-like receptor family pyrin domain containing 3 (NLRP3) expression by corneal epithelium in animals subjected to alkali burn (AB) at 2 days (2 d) and 5 days (5 d) post-injury. (A) Gene expression analysis of NLRP3 messenger mRNA (mRNA) transcript in whole cornea of animals subjected to alkali burn (n = 8 animals/group). Graphs show means ± standard error of the mean (SEM). **** p < 0.0001; (B) Representative merged digital images of laser scanning confocal microscopy of corneas cryosections immunostained for NLRP3 (green) with propidium iodide nuclei counterstaining (DNA in red) in corneas subjected to alkali burn (images are representative of n = 6 animals/group). Scale bar: 50 µm; (C) Representative digital images of western blot of NLRP3 and beta -actin in cornea epithelium of animals subjected to alkali burn for 2 and 5 days. Each lane is a different sample (n = 12 animals/3 samples/group). MW: Molecular weight; UT: untreated control; 2 d and 5 d refer to mice subjected to alkali burn and euthanized 2 or 5 days post-injury, respectively (1 and 2 indicate different samples). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28273882), licensed under a CC-BY license. Not internally tested by R&D Systems.