anti kv1 3 antibody  (Alomone Labs)

Journal: Frontiers in Toxicology

Article Title: Kv1.3 as an upstream regulator of oxidative stress-mediated neuroinflammation following organic dust exposure in murine in vitro , ex vivo , and in vivo models

doi: 10.3389/ftox.2026.1765108

Figure Lengend Snippet: Transcriptomic profiling, qPCR validation, and protein expression analyses reveal microglial responses to ODE exposure in mouse brains. After 5 weeks of exposure to organic dust extract (ODE), brains were collected from male mice (n = 3/group), and the microglial fraction was isolated. Total RNA was extracted from isolated microglia for RNA-seq analysis, and differentially expressed genes (DEGs) were identified based on statistical significance (p ≤ 0.05) and a log 2 fold-change >2. These DEGs are visualized using a volcano plot and bar graph (a,b) . Pathway enrichment analysis using DAVID focused on gene ontology (GO) terms, highlighting significantly enriched pathways related to biological processes (BP) and molecular functions (MF) (c) . Validation of selected DEGs was performed via qPCR analysis. From 40 genes initially selected , six genes demonstrated significant validation (log 2 fold-change >2) compared to controls: Hypoxia-inducible factor 1α (Hif1α), Hypoxia-inducible factor 2α (Hif2α), E2F transcription factor 2 (E2f2), DEAD (Asp-Glu-Ala-Asp) box polypeptide 3, Y-linked (Ddx3y), Potassium voltage-gated channel (Kcna3), and Xylosyltransferase 1 (Xylt1) (d) . Additionally, Western blot analysis was conducted on whole-cell lysates of isolated microglia from freshly dissected mouse brains to assess protein expression levels of Kv1.3 (75 kDa), NOX2 (57 kDa), phosphorylated-p38 MAPK14 (p-p38 MAPK14, 40 kDa), and β-actin (housekeeping protein, 42 kDa). Densitometric quantification revealed significantly increased expression levels of Kv1.3, NOX2, and p-p38 MAPK14 proteins following ODE exposure compared to controls (e–g) . Please refer to for ImageJ based densitometry analysis. Statistical significance is indicated by an asterisk (* for the ODE exposure effect, control vs. ODE). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by one-way ANOVA with Tukey’s multiple comparisons.

Article Snippet: Kv1.3 , APC-101 , Almone labs.

Techniques: Biomarker Discovery, Expressing, Isolation, RNA Sequencing, Western Blot, Control

Journal: Frontiers in Toxicology

Article Title: Kv1.3 as an upstream regulator of oxidative stress-mediated neuroinflammation following organic dust exposure in murine in vitro , ex vivo , and in vivo models

doi: 10.3389/ftox.2026.1765108

Figure Lengend Snippet: Electrophysiological and morphological changes in primary microglia following LPS and ODE stimulation. Primary microglia were either treated with vehicle (DMEM in 2% FBS), LPS (100 ng/mL), or ODE (1% v/v), and whole-cell patch-clamp recordings were performed at 48 h. Light microscopic microphotographs are shown to visualize morphological changes in microglia (a–c) . Kv1.3 and Kir2.1 current density following treatment with DMEM with 2% FBS (control), LPS (100 ng/mL), and ODE (1% w/v) after 48-h are shown as scatterplots from individual cells (d–f) . Representative potassium current recordings from control, LPS or ODE treated microglia (g–i) . The numbers in parentheses show the number of individual cells that were recorded. * signifies differences compared to the unstimulated condition, and the # signifies differences between the ODE and LPS-treated groups. * /# p < 0.05 by unpaired t-test to compare the two groups for statistical significance.

Article Snippet: Kv1.3 , APC-101 , Almone labs.

Techniques: Patch Clamp, Control

Journal: Frontiers in Toxicology

Article Title: Kv1.3 as an upstream regulator of oxidative stress-mediated neuroinflammation following organic dust exposure in murine in vitro , ex vivo , and in vivo models

doi: 10.3389/ftox.2026.1765108

Figure Lengend Snippet: IHC and ICC analysis of Kv1.3 expression in ODE-treated BSCs, BV2 microglia, and primary microglia with PAP-1 treatment effects. Following the treatments , BSCs (a) , BV2 microglia (b) , and primary microglia (referred to as 1°µglia) (c) were immunostained for Kv1.3 (Cy3, red). The antibodies used for immunostaining are detailed in . Nuclei were counterstained with DAPI (blue) to visualize and count the number of cells in each field. To specifically identify microglia expressing Kv1.3 in BSCs, co-staining with Iba1 (a microglia marker) was performed. Images of Iba1, FITC (green) panel (not included in the main manuscript) are provided in . Quantification of IHC/ICC staining for Kv1.3 expression was carried out as described in the Methods section. Compared to the control group, ODE-exposed groups showed a significant increase in Kv1.3 staining intensity in BSCs (b), BV2 microglia (d) , and primary microglia (referred to as 1°µglia) (f) . Notably, PAP-1 treatment significantly reduced Kv1.3 expression in BSCs (a,b) , BV2 microglia (c,d) , and primary microglia (e–f) , as shown in the representative images and quantitative analysis. The sample size included n = 3 mouse pups for BSC and primary microglia, and n = 3 independent cultures for BV2 microglia (biological replicates). Statistical significance is indicated by an * for the ODE exposure effect (control vs. ODE) and # for the PAP-1 treatment effect (ODE vs. ODE + PAP1). The scale bar represents 50 μm * /# p < 0.05, ** /## p < 0.01, *** /### p < 0.001, **** /#### p < 0.0001 by one-way ANOVA with Tukey’s multiple comparisons.

Article Snippet: Kv1.3 , APC-101 , Almone labs.

Techniques: Expressing, Immunostaining, Staining, Marker, Control

Journal: Frontiers in Toxicology

Article Title: Kv1.3 as an upstream regulator of oxidative stress-mediated neuroinflammation following organic dust exposure in murine in vitro , ex vivo , and in vivo models

doi: 10.3389/ftox.2026.1765108

Figure Lengend Snippet: An overview of ODE exposure-induced kcna3 upregulation in primary microglia. ODE exposure mediated an increase in p38 MAPK phosphorylation, leading to upregulated kcna3 expression. This was accompanied by elevated Kv1.3 protein levels in microglia following ODE stimulation. Additionally, ODE exposure triggered the secretion of pro-inflammatory cytokines and nitrite. Treatment with PAP-1 effectively reduced Kv1.3 protein levels, p38 MAPK phosphorylation, and nitrooxidative stress markers. However, PAP-1 treatment did not significantly alter pro-inflammatory cytokine secretion in response to ODE.

Article Snippet: Kv1.3 , APC-101 , Almone labs.

Techniques: Phospho-proteomics, Expressing

Journal: Pflugers Archiv

Article Title: Just a little prick: careful cell contacts enabled by ceramic nanostraws

doi: 10.1007/s00424-026-03150-7

Figure Lengend Snippet: Protein and mRNA expression of typical ion channels in THP-1 cells. Monocytic THP-1 cells have been stimulated with PMA overnight (under exactly the same conditions as for the nanostraw experiments). After that, cells were incubated under normoxia (24 h; NOX) or under hypoxic conditions (HOX: 1% O 2 ) for 2 h, 4 h, and 24 h respectively. ( a ) mRNA expression of TRPM2 , TRPM4 and KCNA3 (encoding Kv1.3) under the given conditions. Whereas TRPM2 showed a significant downregulation after 24 h of hypoxic incubation, TRPM4 mRNA was upregulated. KCNA3 mRNA showed no changes (mean ± SD; n = 9; one way ANOVA plus Dunnett´s multiple comparisons test; ***: p < 0.001, ****: p < 0.0001). ( b ) TRPM4 and Kv1.3 protein are expressed in THP-1 cells under normoxic conditions and can be found in the cytosol and membrane of the cells (400x magnification, scale bar: 20 μm) but did not show altered expression under hypoxia (data not shown). ( c ) TRPM2 protein was cleaved under hypoxic conditions and shorter versions of the protein became detectable after only 2 h of hypoxia. 50 µg of whole cell lysate have been applied to Western blotting. Blot is representative for three independent results. ( d ) Intracellular protein distribution analysis of TRPM2 showed a prominent overall downregulation of TRPM2 after 2 h of hypoxia especially in areas close to the cell membrane. This effect seemed to recover over time (400x magnification, scale bar: 20 μm). Negative control (without primary antibody) did not show staining for TRPM2 at all

Article Snippet: The first antibody (anti-human TRPM2 from rabbit, RRID: AB_1716520, VWR International GmbH, Darmstadt, Germany, #ABNOPAB11990), anti-human TRPM4 from rabbit, (VWR International GmbH, Darmstadt, Germany, #BOSSBS-9051R, RRID: AB_3712801) or anti-human Kv1.3 from rabbit (#APC-101, RRID: AB_2040149, Alomone labs, Jerusalem, Israel) was added at a concentration of 1:100 in 3% BSA-PBS for 2 h. Cells were washed with PBS before the second antibody (goat anti-rabbit Alexa Fluor488 (#A-11008; RRID: AB_143165) for TRPM2 and goat anti-rabbit Alexa Fluor568 (#A-11011, RRID: AB_143157) for TRPM4 and Kv1.3, both from Life Technologies, Darmstadt, Germany and used in a dilution of 1:400 in PBS) was added for additional 90 min. Cover glasses were fixed with fluorescence mounting medium (#S3023, Agilent, Santa Clara, United States of America) on the slides, and cells were analyzed the following day with a fluorescence microscope (AxioVert with Axiocam 305; Zeiss, Oberkochen, Germany).

Techniques: Expressing, Incubation, Membrane, Western Blot, Negative Control, Staining

Journal: bioRxiv

Article Title: Kv1.3 palmitoylation regulates spatial distribution and channel removal from the immunological synapse

doi: 10.64898/2026.01.19.700329

Figure Lengend Snippet: Palmitoylation facilitates the ubiquitination and central accumulation of the channel at the immunological synapse. (A) Representative TIRF images of CD4 + T cells electroporated with WT or Cys less Kv1.3 YFP to form synapses on the SLB. The ICAM1 ring indicates the formation of a synapse. The merged image shows Kv1.3 (green) and ICAM1 (blue) expression. IRM, interference reflection microscopy. Scale bars represent 5 μm. (B) Quantification of Kv1.3 intensity at the immunological synapse normalized to the average of the WT for each donor. Data are presented as the means ± SEs of n > 300 cells from 3 independent blood donors. **p < 0.01 by Student’s t test. (C) ROIs of each supramolecular activation complex (SMAC) in (A). IRM images were used to define the total cell contact area. The ICAM1 ring indicates the pSMAC. IRM regions outside or inside the ICAM1 ring were classified as dSMAC or cSMAC, respectively. (D) Kv1.3 intensity was quantified within each ROI corresponding to the cSMAC, pSMAC, dSMAC, and the entire synapse. The percentage of Kv1.3 intensity in each SMAC was calculated with respect to the total number of synapses. Data are presented as the means ± SEs of n > 50 cells from 3 independent blood donors. *p<0.05, ****p < 0.0001 by Student’s t test. (E) A PLA was performed using anti-ubiquitin and anti-Kv1.3 antibodies to detect ubiquitinated Kv1.3. Orthogonal views from a representative confocal image of ubiquitinated Kv1.3 (PLA signal, magenta) in a synapse conjugate between a human CD4 + T cell and a Raji B cell. CD19 (gray) was used as a B-cell-specific marker, and phalloidin (cyan) was used to stain the actin filaments. The PLA signal detected within the synaptic contact is located at the center of the synapse (XZ plane). (F) Ubiquitination assay in HEK293 cells transfected with either WT or Cys less Kv1.3 YFP. Cells were incubated in the absence (-) or presence (+) of phorbol 12-myristate 13-acetate (PMA) for 30 min to induce protein internalization as a positive control. Cell lysates were immunoprecipitated (IP) for Kv1.3 and immunoblotted (IB) for both Kv1.3 and ubiquitin (Ubq). SM: starting materials, IP: immunoprecipitation, IP - : negative control in the absence of antibody. (G) Quantification of channel ubiquitination. The data are presented as the means ± SEs of at least 2 independent experiments. **p < 0.01 by Student’s t test.

Article Snippet: Next, the cells were labeled with primary anti-CD3 (Alexa Fluor 647; Biolegend), anti-CD19 (Brilliant Violet 421; Biolegend), anti-ZDHHC21 (NSJ Bioreagents) and anti-Kv1.3 (FITC; Alomone) antibodies in 1% BSA for 1 h at RT.

Techniques: Ubiquitin Proteomics, Expressing, Microscopy, Activation Assay, Marker, Staining, Transfection, Incubation, Positive Control, Immunoprecipitation, Negative Control