rabbit anti trpv4 polyclonal antibody  (Alomone Labs)

Journal: Toxics

Article Title: The Synergistic Effects of Fine Particulate Matter and High Humidity on Allergic Asthma: An Association with TRPV4/MAPK Pathway Activation

doi: 10.3390/toxics14030219

Figure Lengend Snippet: The experimental protocol. ( A ) Schematic timeline of sensitisation, challenge and exposure for the 12 experimental groups; ( B ) Schematic overview of the in vivo exposure system; ( C ) Chemical structure of the TRPV4 antagonist HC-067047.

Article Snippet: To minimize non-specific binding, slides were incubated with 3% bovine serum albumin (BSA) in PBS containing 0.05% ( v / v ) Tween-20 (PBST) at room temperature for 1 h, followed by overnight exposure to rabbit anti-TRPV4 polyclonal antibody (Alomone Labs, Jerusalem, Israel; 1:100) at 4 °C.

Techniques: In Vivo

Journal: Toxics

Article Title: The Synergistic Effects of Fine Particulate Matter and High Humidity on Allergic Asthma: An Association with TRPV4/MAPK Pathway Activation

doi: 10.3390/toxics14030219

Figure Lengend Snippet: The impact of exposure to PM 2.5 and different relative humidities on the TRPV4 ion channel in the lung tissue. ( A ) Immunohistochemistry for TRPV4 in the lung tissue (original magnification, 200×). ( B ) The average optical density of TRPV4 in the lung tissue. * p < 0.05, ** p < 0.01: a significant difference compared with the saline group. ns p > 0.05, ## p < 0.01: different exposure groups compared with the OVA group. †† p < 0.01: the blocking groups compared with the corresponding exposure groups ( n = 7).

Article Snippet: To minimize non-specific binding, slides were incubated with 3% bovine serum albumin (BSA) in PBS containing 0.05% ( v / v ) Tween-20 (PBST) at room temperature for 1 h, followed by overnight exposure to rabbit anti-TRPV4 polyclonal antibody (Alomone Labs, Jerusalem, Israel; 1:100) at 4 °C.

Techniques: Immunohistochemistry, Saline, Blocking Assay

Journal: Acta Neuropathologica Communications

Article Title: Blocking axon-glial mechanotransduction to prevent concussive brain injury

doi: 10.1186/s40478-025-02117-6

Figure Lengend Snippet: Protective effects of GSK219 are absent in TRPV4 −/− mice after head impact in CHIMERA. ( A ) Low-mag (left) and confocal (right) images from the cortex of TRPV4 −/− mice received no impact (Sham, top), or 0 h after CHIMERA (0.9 J) with vehicle (middle) or GSK219 pretreatment (bottom). YFP signals are inverted in gray-scale images and green in confocal images. Scale bars, 200 μm in left panels, and 20 μm in right panels. ( B ) Summary of the effects of TRPV4 KO and GSK219 pretreatment on axonal varicosity formation at 0 h after CHIMERA. ( C ) Summary of the effects of TRPV4 KO and GSK219 pretreatment on dMBP staining intensities in the cortex 24 h after CHIMERA. In ( B ) and ( C ), One-way ANOVA followed by Dunnett’s test: *** p < 0.001. Mouse numbers: 4 (Sham), 5 (Veh 0 h), and 4 (GSK 0 h). 1–3 images from each mouse are included. ( D ) Protein changes in the brains of TRPV4 −/− mice versus age- and sex-matched WT mice revealed by genome-wide proteomics with mass spectrometry analysis. ( E ) Upregulated proteins in the TRPV4 −/− mouse brain in GO subgroups based on subcellular components. ( F ) Down-regulated proteins in the TRPV4 −/− mouse brain in GO subgroups. ( G ) The upregulated ion channel proteins versus unchanged Cav channels. Unpaired t-test; Kcnj3: * p = 0.0075; Cacna1a: * p = 0.0024; Cacna1e: * p = 0.011; Mcu: * p = 0.0049 ; Grik2: * p = 0.038; Ryr2: * p = 0.033; Scn2a: * p = 0.027. ( H ) TRPV4 deletion was verified by Western blotting with WT and TRPV4 −/− brains. ( I ) Western blots of WT and TRPV4 −/− brains using antibodies against 3 Cav channel proteins. Numbers on the right, molecular weights in kDa. ( J ) Quantification summary of Western blot results. Unpaired t-test: * p < 0.05

Article Snippet: Antibodies used in this study include a rabbit polyclonal anti-degraded MBP antibody that recognizes damaged MBP proteins (1:2000, anti-dMBP; Catalog #: AB5864; Chemicon), a rat monoclonal anti-MBP antibody (1:200; Catalog #: ab7349; abcam), a rabbit polyclonal anti-TRPV4 antibody (1:200; Catalog #: LS-A8583; LifeSpan BioSciences, Seattle, Washington, USA), rabbit polyclonal anti-Cav2.1, anti-Cav2.2 and anti-Cav2.3 antibodies (Catalog #s: ACC-001, ACC-002 and ACC-006; Alomone Labs, Jerusalem, Israel), a goat polyclonal anti-GFAP antibody (1:200; Catolog #: Ab53554; Abcam Inc. Waltham, MA, USA), and a rat monoclonal anti-CD68 antibody (1:200; Catalog #: MCA1957GA; Bio-Rad; Hercules, CA, USA), as well as Cy3 and Cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Techniques: Staining, Genome Wide, Mass Spectrometry, Western Blot

Journal: Acta Neuropathologica Communications

Article Title: Blocking axon-glial mechanotransduction to prevent concussive brain injury

doi: 10.1186/s40478-025-02117-6

Figure Lengend Snippet: Acute deletion of neuronal TRPV4 prevents CHIMERA-induced axonal varicosity formation and adjacent glial changes. ( A ) Diagram of injecting AAV9-hSyn-Cre-dTomato into the right cortex of mice. Adult WT and TRPV4 fl/fl mice were used in CHIMERA about 1 month after the viral injection. A low-mag image (bottom) shows the injection sites with dTomato fluorescence in green and Hoechst in blue. ( B ) Low-mag gray-scale images with dTomato signals inverted of the cortex and EC from a WT (top) or a TRPV4 fl/fl (bottom) mouse injected with AAV9:Syn-Cre; dTomato at 0 h after CHIMERA. ( C ) Confocal images from the EC of the mice in ( B ). The dTomato signals are in green and Hoechst in blue. ( D ) Summary of the percentage of dTomato + axons with varicosities in white matter (CC and EC) and gray matter (cortical layers 1–6) from WT (black) and TRPV4 fl/fl (red) mice 0–24 h after CHIMERA, compared to Sham. n = 7 for all conditions. ( E ) Confocal images and quantification of TRPV4 protein (red) knockdown via the infection by AAV9-Syn-Cre-dTomato (green) in the cortex of TRPV4 fl/fl but not WT mice. Unpaired t-test: **, p < 0.01. n = 7 for both conditions. ( F ) Confocal images from the Ipsilateral and Contralateral sides of the cortex from WT (top) or TRPV4 fl/fl (bottom) mice 24 h after CHIMERA. The dTomato signals are in green and dMBP signals are in red. ( G ) Summary of dMBP staining intensity from the mouse cortex under different conditions. ( H ) Summary of CD68 + cell density in the cortex from WT or TRPV4 fl/fl (Flox) mice with AAV injection 24 h after CHIMERA. n = 10 in all groups. Unpaired t-test: **, p < 0.01; ***, p < 0.001. Scale bars, 160 μm in ( B ); 40 μm in ( C ), ( E ), and ( F )

Article Snippet: Antibodies used in this study include a rabbit polyclonal anti-degraded MBP antibody that recognizes damaged MBP proteins (1:2000, anti-dMBP; Catalog #: AB5864; Chemicon), a rat monoclonal anti-MBP antibody (1:200; Catalog #: ab7349; abcam), a rabbit polyclonal anti-TRPV4 antibody (1:200; Catalog #: LS-A8583; LifeSpan BioSciences, Seattle, Washington, USA), rabbit polyclonal anti-Cav2.1, anti-Cav2.2 and anti-Cav2.3 antibodies (Catalog #s: ACC-001, ACC-002 and ACC-006; Alomone Labs, Jerusalem, Israel), a goat polyclonal anti-GFAP antibody (1:200; Catolog #: Ab53554; Abcam Inc. Waltham, MA, USA), and a rat monoclonal anti-CD68 antibody (1:200; Catalog #: MCA1957GA; Bio-Rad; Hercules, CA, USA), as well as Cy3 and Cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Techniques: Injection, Fluorescence, Knockdown, Infection, Staining

Journal: Acta Neuropathologica Communications

Article Title: Blocking axon-glial mechanotransduction to prevent concussive brain injury

doi: 10.1186/s40478-025-02117-6

Figure Lengend Snippet: GSK279 markedly inhibits CHIMERA-induced axon-glial changes in WT but not TRPV4 −/− mice. ( A ) Structural diagrams of GSK219 and GSK279. ( B ) Diagram for the CHIMERA experiment with GSK279 pretreatment. WT; Thy1-YFP or TRPV4 −/− ;Thy1-YFP mice (3–4 months old) were injected via tail vein with GSK279 (18 µg/kg) or vehicle (as control) once 1.5 h before CHIMERA (0.9 J). Mice were perfused and fixed either immediately (0 h) or 24 h after head impact. ( C ) Summary of YFP + axons with varicosities in EC 0 h after CHIMERA with vehicle or GSK279 pretreatment. The green line, the basal level in Sham. ( D ) Confocal images of the EC of WT (left) and TRPV4 −/− (right) mice 24 h after CHIMERA with the pretreatment of vehicle (top) or GSK279 (bottom). The CD68 staining signals are inverted in grayscale images (right) and red in merged images (left), YFP in green, and Hoechst in blue. ( E ) Summary of YFP + axons with varicosities in EC 24 h after CHIMERA. ( F ) Summary of CD68 + cell density in the cortex 24 h after CHIMERA. ( G ) The dMBP staining signals (inverted in gray-scale images (right) and in red in merged images (left) in the cortex of WT mice 24 h after CHIMERA with vehicle (top) or GSP279 (middle) pretreatment. The summary is at the bottom. ( H ) The dMBP staining signals in the cortex of TRPV4 −/− mice 24 h after CHIMERA with vehicle or GSK279 pretreatment. Mouse numbers, n = 3 for all groups. 1–3 images from each mouse are included. Unpaired t-test: ***, p < 0.001. Scale bars, 20 μm in ( D ), ( G ), and ( H )

Article Snippet: Antibodies used in this study include a rabbit polyclonal anti-degraded MBP antibody that recognizes damaged MBP proteins (1:2000, anti-dMBP; Catalog #: AB5864; Chemicon), a rat monoclonal anti-MBP antibody (1:200; Catalog #: ab7349; abcam), a rabbit polyclonal anti-TRPV4 antibody (1:200; Catalog #: LS-A8583; LifeSpan BioSciences, Seattle, Washington, USA), rabbit polyclonal anti-Cav2.1, anti-Cav2.2 and anti-Cav2.3 antibodies (Catalog #s: ACC-001, ACC-002 and ACC-006; Alomone Labs, Jerusalem, Israel), a goat polyclonal anti-GFAP antibody (1:200; Catolog #: Ab53554; Abcam Inc. Waltham, MA, USA), and a rat monoclonal anti-CD68 antibody (1:200; Catalog #: MCA1957GA; Bio-Rad; Hercules, CA, USA), as well as Cy3 and Cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Techniques: Injection, Control, Staining

Journal: Acta Neuropathologica Communications

Article Title: Blocking axon-glial mechanotransduction to prevent concussive brain injury

doi: 10.1186/s40478-025-02117-6

Figure Lengend Snippet: Gabapentin markedly inhibits CHIMERA-induced axon-glial changes in TRPV4 −/− mice. ( A ) Diagram for the CHIMERA experiment with gabapentin pretreatment. WT; Thy1-YFP or TRPV4 −/− ;Thy1-YFP mice were injected via I.P. with gabapentin (50 mg/kg) or vehicle (as control) once 1.5 h before CHIMERA (0.9 J). Mice were perfused and fixed either immediately (0 h) or 24 h after head impact. ( B ) Summary of YFP + axons with varicosities in EC 0 h after CHIMERA with vehicle or gabapentin pretreatment. The green line, the basal level in Sham. ( C ) Summary of YFP + axons with varicosities in EC 24 h after CHIMERA. ( D ) Confocal images of the EC of WT (left) and TRPV4 −/− (right) mice 24 h after CHIMERA with the pretreatment of vehicle (Veh; top) or gabapentin (Gab; bottom). The CD68 staining signals are inverted in grayscale images (right) and in red in merged images (left), YFP in green, and Hoechst in blue. ( E ) Summary of CD68 + cell density in the cortex 24 h after CHIMERA. ( F ) The dMBP staining signals in the cortex of WT (left) or TRPV4 −/− (right) mice 24 h after CHIMERA. ( G ) Summary of dMBP signals in the cortex 24 h after CHIMERA. Mouse numbers, n = 3 for all groups. 1–3 images from each mouse are included. Unpaired t-test: **, p < 0.01; ***, p < 0.001. Scale bars, 25 μm in ( D ) and ( F )

Article Snippet: Antibodies used in this study include a rabbit polyclonal anti-degraded MBP antibody that recognizes damaged MBP proteins (1:2000, anti-dMBP; Catalog #: AB5864; Chemicon), a rat monoclonal anti-MBP antibody (1:200; Catalog #: ab7349; abcam), a rabbit polyclonal anti-TRPV4 antibody (1:200; Catalog #: LS-A8583; LifeSpan BioSciences, Seattle, Washington, USA), rabbit polyclonal anti-Cav2.1, anti-Cav2.2 and anti-Cav2.3 antibodies (Catalog #s: ACC-001, ACC-002 and ACC-006; Alomone Labs, Jerusalem, Israel), a goat polyclonal anti-GFAP antibody (1:200; Catolog #: Ab53554; Abcam Inc. Waltham, MA, USA), and a rat monoclonal anti-CD68 antibody (1:200; Catalog #: MCA1957GA; Bio-Rad; Hercules, CA, USA), as well as Cy3 and Cy5-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Techniques: Injection, Control, Staining

Journal: The Journal of Physiological Sciences : JPS

Article Title: Calcium response via CRAC channels in human synovial cells induced by shear stress in rheumatoid arthritis

doi: 10.1016/j.jphyss.2025.100013

Figure Lengend Snippet: Immunofluorescence staining, differential interference contrast (DIC) microscopy images, and immunofluorescence intensity of the normal and RA FLS cells, Immunofluorescence staining with anti-Ca 2+ release-activated channel regulator 2 A (CRACR2A) antibody (green, Alexa) and DIC images of the normal (A) and RA (B) FLS cells. Immunofluorescence staining with the anti-transient receptor potential vanilloid 4 (TRPV4) antibody (red, Alexa) and DIC images of the normal (C) and RA (D) FLS cells. All fluorescence and DIC images were obtained at 20 × magnification using an Olympus FV1000 Confocal Laser Microscope. Scale bar = 100 µm. The mean immunofluorescence intensity of the anti-CRACR2A antibody staining was significantly lower in the normal FLS cells (4.08 ± 0.8 AU) than in the RA FLS cells (20.3 ± 6.9 AU) (p < 0.01). The mean immunofluorescence intensities produced by the anti-TRPV4 antibody staining were not significantly different between the normal (30.3 ± 6.2 AU) and RA (29.7 ± 6.2 AU) FLS cells (E). n.s.: not significant, * *: p < 0.01 , compared between the normal and RA FLS cells.

Article Snippet: After fixation, the cultures were washed twice with PBS and blocked with 0.2 % bovine serum in PBS for 30 min. Primary antibodies were diluted (1:250 for rabbit polyclonal anti-CRACR2A antibody [66787–1-IG, Proteintech, IL, USA] and 1:250 for rabbit polyclonal anti-TRPV4 antibody [ab191580, Alomone Labs, Jerusalem, Israel]) and incubated for 1 h at 37.5 °C.

Techniques: Immunofluorescence, Staining, Microscopy, Fluorescence, Produced

Journal: The Journal of Physiological Sciences : JPS

Article Title: Calcium response via CRAC channels in human synovial cells induced by shear stress in rheumatoid arthritis

doi: 10.1016/j.jphyss.2025.100013

Figure Lengend Snippet: Expression levels of ORAI1, stromal interaction molecule 1 (STIM1), CRACR2A, and TRPV4 in the normal and RA FLS cells Western blotting of ORAI1, STIM1, CRACR2A, and TRPV4 expression in the normal and RA FLS cells. (A) The internal controls and representative western blots are shown. Subsequently, 10 or 30 μg of protein per lane was loaded onto the gel. Next, the membranes were probed with specific antibodies against Orai1, STIM1, CRACR2A, or TRPV4 with appropriate secondary antibodies. Protein bands were detected using C-DiGit (LI-COR#3600–00), and the results were analyzed using the ImageJ software (B). The protein amounts were normalized to β-actin amounts to indicate the protein expression ratios. * * and † † represent significant differences from normal FLS, respectively, at p < 0.01.

Article Snippet: After fixation, the cultures were washed twice with PBS and blocked with 0.2 % bovine serum in PBS for 30 min. Primary antibodies were diluted (1:250 for rabbit polyclonal anti-CRACR2A antibody [66787–1-IG, Proteintech, IL, USA] and 1:250 for rabbit polyclonal anti-TRPV4 antibody [ab191580, Alomone Labs, Jerusalem, Israel]) and incubated for 1 h at 37.5 °C.

Techniques: Expressing, Western Blot, Software

Journal: The Journal of Physiological Sciences : JPS

Article Title: Calcium response via CRAC channels in human synovial cells induced by shear stress in rheumatoid arthritis

doi: 10.1016/j.jphyss.2025.100013

Figure Lengend Snippet: Ca 2+ influx from the CRAC and TRPV4 of synovial cells in response to SS, When the Ca 2+ in the endoplasmic reticulum was depleted by SS, CRACR2A directly interacted with the cytoplasmic regions of ORAI1 and STIM1 to form a ternary complex, resulting in the intracellular Ca 2+ uptake from extracellular sources. In the RA FLS cells, STIM1 and CRACR2A were overexpressed compared with the expression in the normal FLS cells, and the regulation of intracellular Ca 2+ concentration significantly depended on CRAC. SS also induced Ca 2+ uptake from the extracellular into the intracellular site via TRPV4 and/or other mechanosensitive Ca 2+ channels. However, there were no significant difference in the expression level of TRPV4 between the normal and RA FLS cells, indicating that TRPV4 exerted the same influence on the regulation of intracellular Ca 2+ concentration in both FLS cell types.

Article Snippet: After fixation, the cultures were washed twice with PBS and blocked with 0.2 % bovine serum in PBS for 30 min. Primary antibodies were diluted (1:250 for rabbit polyclonal anti-CRACR2A antibody [66787–1-IG, Proteintech, IL, USA] and 1:250 for rabbit polyclonal anti-TRPV4 antibody [ab191580, Alomone Labs, Jerusalem, Israel]) and incubated for 1 h at 37.5 °C.

Techniques: Expressing, Concentration Assay

Journal: Frontiers in Medicine

Article Title: The co-expression of the depolarizing and hyperpolarizing mechanosensitive ion channels in mammalian retinal neurons

doi: 10.3389/fmed.2024.1463898

Figure Lengend Snippet: Antibodies.

Article Snippet: Polyclonal rabbit anti-TRPV4 (LS-C135, 1: 200; LS-A8583 1:200 and LS-C94498 1: 100) was purchased from LifeSpan Biosciences, Inc. (Seatle, WA, USA).

Techniques: Mutagenesis, Recombinant, Negative Control, Transduction, Western Blot, Knock-Out

Journal: Frontiers in Medicine

Article Title: The co-expression of the depolarizing and hyperpolarizing mechanosensitive ion channels in mammalian retinal neurons

doi: 10.3389/fmed.2024.1463898

Figure Lengend Snippet: The expression of TRAAK and TRPV4 in the mouse retina. (A,B) Retinal slices were retrogradely labeled for ganglion cells (GCs) by neurobiotin (NB) and stained for TRPV4 (green) and TRAAK ( A , pink) or glutamine synthetase (GS, blue, B ). TRPV4 signals are heavier in the GCL. (A) The image was inverted and displaced on a white background. TRAAK is heavily expressed in Müller cells and weakly expressed in OSL and OPL. (B) Axons and somas of GCs are brightly positive for TRPV4. TRPV4 is present in Müller cells (MC). Some somas of putative cone BCs in the second soma row of the INL contain no TRPV4 signals (open arrow, inset b). (C,D) Retinal slices from wild-type (w.t.) and TRPV4 transgenic mice (TRPV4−/−) were labeled for TRPV4 (green) and PKCa (red). (C) Some TRPV4 puncta are present in rod BCs (asterisks) and cone BCs (triangles). (D) TRPV4 signal is nearly absent in TRPV4 transgenic mice. OSL: outer segment layer; ISL: inner segment layer; OPL: outer plexiform layer; INL: inner nuclear layer; BCL-bipolar cell layer; ACL-amacrine cell layer; IPL-inner plexiform layer; GCL-ganglion cell layer; NFL-nerve fiber layer. The scale bar is 5 μm for C and 20 μm for others.

Article Snippet: Polyclonal rabbit anti-TRPV4 (LS-C135, 1: 200; LS-A8583 1:200 and LS-C94498 1: 100) was purchased from LifeSpan Biosciences, Inc. (Seatle, WA, USA).

Techniques: Expressing, Labeling, Staining, Transgenic Assay

Journal: Frontiers in Medicine

Article Title: The co-expression of the depolarizing and hyperpolarizing mechanosensitive ion channels in mammalian retinal neurons

doi: 10.3389/fmed.2024.1463898

Figure Lengend Snippet: TRPV4 mediates excitatory visual noises and dysfunction of mouse RGCs. RGCs were recorded under loose patch mode (A,B) and whole-cell voltage-clamp mode at various holding potentials (D–G) . (A,B,F,G) TRPV4 agonists enhance spontaneous action potentials ( A,B , arrow) and the spontaneous postsynaptic currents (arrow) and light-evoked excitatory currents (F,G) , reducing the visual signal reliability [VSR = 1-noise (N)/signal (S)] by ~50% (C) . (D,E) Confocal images of a recorded RGC filled with Lucifer yellow ( D : The x-y view; E : The y-z view revealing the dendritic tree and axon of the cell).

Article Snippet: Polyclonal rabbit anti-TRPV4 (LS-C135, 1: 200; LS-A8583 1:200 and LS-C94498 1: 100) was purchased from LifeSpan Biosciences, Inc. (Seatle, WA, USA).

Techniques:

Journal: Frontiers in Medicine

Article Title: The co-expression of the depolarizing and hyperpolarizing mechanosensitive ion channels in mammalian retinal neurons

doi: 10.3389/fmed.2024.1463898

Figure Lengend Snippet: The activation of MSCs induces membrane depolarization, ATP depletion, and dysfunction of mouse RGCs. RGCs are recorded under current-clamp (A,B) and voltage-clamp (C) modes. (A,B) Activating MSCs by low osmolarity (Osm) and TRPV4 agonists 4aPDD and GSK101 (GSK) depolarizes MP of RGCs to −40 to −50 mV and elicits spontaneous firing of action potentials (A) , which raises the ATP consume close to the peak level and predicts ATP depletion. (C) The threshold of NaVs in mouse RGCs is close to −50 mV and sensitive to TTX. The duration of the sodium current is ~2 ms. (D) Blue curve: the standard ATP consumption for RGCs to maintain RP was plotted per . TRPV4 agonists raised the ATP consumption (green dot) near the inactivation level of half NaVs (purple dot). Vh: holding potential. MP: membrane potential. RP: resting potential. NaV: voltage-gated sodium channel. RR: ruthenium red, nonspecific blocker of TRPs.

Article Snippet: Polyclonal rabbit anti-TRPV4 (LS-C135, 1: 200; LS-A8583 1:200 and LS-C94498 1: 100) was purchased from LifeSpan Biosciences, Inc. (Seatle, WA, USA).

Techniques: Activation Assay, Membrane