Human Pax2 Antibody

Detection of Mouse Pax2 by Western Blot. Western blot shows lysates of mouse fetal kidney. PVDF Membrane was probed with 1 µg/mL of Goat Anti-Human Pax2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3364) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for Pax2 at approximately 47 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Pax2 in BG01V Human Embyonic Stem Cells. Pax2 was detected in immersion fixed BG01V human embryonic stem cells differentiated into the early otic lineage using Goat Anti-Human Pax2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3364) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Pax2 in Human Kidney. Pax2 was detected in immersion fixed paraffin-embedded sections of human kidney using Goat Anti-Human Pax2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3364) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei in convoluted tubules. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Rat Human Pax2 Antibody by Immunohistochemistry Ablation of AAV-PdynP+ spinal dorsal horn (SDH) neurons does not affect behavioral responses evoked by the optical stimulation of A beta fibers, nociceptive mechanical force, and heat in rats. (A) Enhanced green fluorescent protein (EGFP) expression (green) in the L4 SDH at 4 weeks after intra-SDH microinjection of AAV-PdynP-DTR-EGFP (indicated by an upper schematic illustration). Colocalization of EGFP and PAX2 (red) in the SDH (indicated by arrowheads). Scale bar, 100 μm. The percentage of co-expressing neurons was quantified (n = 3 rats tested, 94 total EGFP+ cells quantified, and 339 total PAX2+ cells quantified). (B) Diphtheria toxin receptors (DTR) immunofluorescence in the SDH after administration of diphtheria toxin (DTX: 50 μg/kg, i.p., two injection 72 h apart) or PBS. Scale bar, 200 μm. (C) Immunohistochemical analysis and quantification of PAX2+ neurons in the superficial laminae of the ipsilateral and contralateral SDH at 4 weeks after DTX injection (n = 6 rats tested, PAX2+ neurons: 992 [ipsi] and 1574 [contra] cells quantified). Scale bar, 50 μm. ****P < 0.0001, Unpaired t-test. (D) Paw withdrawal behaviors by light (score), von Frey filaments (threshold), and heat (latency) at 4 weeks after DTX or PBS injection (n = 7–9 rats). Data shown as mean ± SEM. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35813063), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Rat Human Pax2 Antibody by Immunohistochemistry Ablation of AAV-PdynP+ spinal dorsal horn (SDH) neurons does not affect behavioral responses evoked by the optical stimulation of A beta fibers, nociceptive mechanical force, and heat in rats. (A) Enhanced green fluorescent protein (EGFP) expression (green) in the L4 SDH at 4 weeks after intra-SDH microinjection of AAV-PdynP-DTR-EGFP (indicated by an upper schematic illustration). Colocalization of EGFP and PAX2 (red) in the SDH (indicated by arrowheads). Scale bar, 100 μm. The percentage of co-expressing neurons was quantified (n = 3 rats tested, 94 total EGFP+ cells quantified, and 339 total PAX2+ cells quantified). (B) Diphtheria toxin receptors (DTR) immunofluorescence in the SDH after administration of diphtheria toxin (DTX: 50 μg/kg, i.p., two injection 72 h apart) or PBS. Scale bar, 200 μm. (C) Immunohistochemical analysis and quantification of PAX2+ neurons in the superficial laminae of the ipsilateral and contralateral SDH at 4 weeks after DTX injection (n = 6 rats tested, PAX2+ neurons: 992 [ipsi] and 1574 [contra] cells quantified). Scale bar, 50 μm. ****P < 0.0001, Unpaired t-test. (D) Paw withdrawal behaviors by light (score), von Frey filaments (threshold), and heat (latency) at 4 weeks after DTX or PBS injection (n = 7–9 rats). Data shown as mean ± SEM. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35813063), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Rat Human Pax2 Antibody by Immunohistochemistry AAV-PdynP injected into rat spinal dorsal horn (SDH) captures a subset of inhibitory neurons. (A) tdTomato (tdT) expression in the fourth lumbar (L4) SDH at 4 weeks after microinjection of AAV-PdynP-tdT. Schematic illustration indicates intra-SDH microinjection of the AAV vector. Scale bar, 200 μm. (B) Immunolabeling of tdT+ cells (red) with lamina-selective markers (green, NK1R; blue, IB4) in the L4 SDH. Scale bar, 200 μm. Quantification of the distribution of tdT+ cells (n = 3 rats tested, 272 total tdT+ cells quantified). (C) Immunofluorescence of prodynorphin (PDYN; green) in tdT+ SDH neurons (red). Scale bar, 200 μm (above), 20 μm (below). Arrowheads indicate tdT+ PDYN+ neurons. The percentage of tdT+ PDYN+ neurons per total tdT+ neurons was quantified (n = 4 rats tested, 1,281 total tdT+ cells quantified). (D) No colocalization of tdT (red) with NK1R (green). Scale bar, 50 μm. The percentage of co-expressing neurons was quantified (n = 3 rats tested, 187 total tdT+ cells quantified). (E) Firing patterns of tdT+ SDH neurons, and the percentage of neurons displaying each pattern. APs were evoked by current injection (80 pA). (F) Colocalization of tdT (red) with PAX2 (blue) and VGLUT2 (green) in the superficial laminae. Arrowheads and arrows indicate tdT+PAX2+ and tdT+VGLUT2+ neurons, respectively. Scale bar, 50 μm. The percentage of co-expressing neurons was quantified (n = 4 rats tested, 456 total tdT+ cells quantified). Data shown as mean ± SEM. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35813063), licensed under a CC-BY license. Not internally tested by R&D Systems.