rabbit anti trpv3  (Boster Bio)

Journal: Communications biology

Article Title: TRPV3-ANO1 interaction positively regulates wound healing in keratinocytes.

doi: 10.1038/s42003-023-04482-1

Figure Lengend Snippet: Fig. 1 TRP channels and ANOs expression in NHEKs. a RT-PCR of TRPs and ANOs in NHEKs. Uncropped gel images are shown in Suppl. Fig. 11. b Western blot of ANO1 in NHEKs. The predicted band size of ANO1 is 114 kDa. c, d Calcium imaging in NHEKs. Cam Camphor, a TRPV3 agonist; GSK GSK1016790A, a TRPV4 agonist; Ca2+ (−) Calcium-free bath solution. e Comparison of current–voltage relationships. The red line indicates current–voltage relationship of basal current in NHEKs using a standard bath and pipette solution. The blue line indicates current–voltage relationship in TRPV6-expressing HEK293T cells using a standard bath solution and NMDG-Cl pipette solution. The holding potential was −60 mV and ramp-pulses were applied from −100 to +100 mV for 300 ms duration every 5 s.

Article Snippet: The following antibodies were used: rabbit anti-ANO1 antibody (Abcam, ab53213, 1:5 for Western blotting), (Abcam, ab53212, 1:100 for immunoprecipitation), rabbit anti-phospho-ERK (extracellular signal-related kinase) antibody (Cell Signaling Technology, #4370, 1:1000), rabbit anti-phosphop38 antibody (Cell Signaling Technology, #4511, 1:1000), rabbit anti-phospho-JNK (c-Jun N-terminal Kinase) antibody (Cell Signaling Technology, #4668, 1:1000), rabbit anti-ERK antibody (Cell Signaling Technology, #4695, 1:1000), rabbit antip38 antibody (Cell Signaling Technology, #8690, 1:1000), rabbit anti-JNK antibody (Cell Signaling Technology, #9252, 1:1000), mouse anti-β-actin antibody (Abcam, ab6276, 1:2500), rabbit anti-TRPV3 antibody (Cell Signaling Technology, #3484, 1:1000 for Western blotting; 1:50 for immunoprecipitation), anti-rabbit-HRP antibody (Cell Signaling Technology, #7074, 1:2000) and anti-mouse-HRP antibody (Cell Signaling Technology, #7076, 1:2000).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Imaging, Comparison, Transferring

Journal: Communications biology

Article Title: TRPV3-ANO1 interaction positively regulates wound healing in keratinocytes.

doi: 10.1038/s42003-023-04482-1

Figure Lengend Snippet: Fig. 2 TRPV3 and ANO1 interaction in HEK293T cells. a A representative trace of the camphor-induced currents in HEK293T cells expressing both hTRPV3 and hANO1, hANO1 alone or hTRPV3 alone. All data were collected using an NMDG-Cl bath and pipette solutions. Free calcium in the pipette solution was 100 nM. The holding potential was −60 mV and ramp-pulses were applied from −100 to +100 mV for 300 ms duration every 5 s. b Comparison of peak currents of (A) at −60 mV (means ± SEM, N = 6). Statistical significance was determined with a Mann–Whitney test. c Immunoprecipitation of ANO1 or TRPV3 and Western blot of TRPV3 in HEK293T cells transfected with TRPV3 and ANO1 cDNAs, Ano1 alone, TRPV4 alone or pcDNA3.1. Uncropped gel images are shown in Suppl. Fig. 11.

Article Snippet: The following antibodies were used: rabbit anti-ANO1 antibody (Abcam, ab53213, 1:5 for Western blotting), (Abcam, ab53212, 1:100 for immunoprecipitation), rabbit anti-phospho-ERK (extracellular signal-related kinase) antibody (Cell Signaling Technology, #4370, 1:1000), rabbit anti-phosphop38 antibody (Cell Signaling Technology, #4511, 1:1000), rabbit anti-phospho-JNK (c-Jun N-terminal Kinase) antibody (Cell Signaling Technology, #4668, 1:1000), rabbit anti-ERK antibody (Cell Signaling Technology, #4695, 1:1000), rabbit antip38 antibody (Cell Signaling Technology, #8690, 1:1000), rabbit anti-JNK antibody (Cell Signaling Technology, #9252, 1:1000), mouse anti-β-actin antibody (Abcam, ab6276, 1:2500), rabbit anti-TRPV3 antibody (Cell Signaling Technology, #3484, 1:1000 for Western blotting; 1:50 for immunoprecipitation), anti-rabbit-HRP antibody (Cell Signaling Technology, #7074, 1:2000) and anti-mouse-HRP antibody (Cell Signaling Technology, #7076, 1:2000).

Techniques: Expressing, Transferring, Comparison, MANN-WHITNEY, Immunoprecipitation, Western Blot, Transfection

Journal: Communications biology

Article Title: TRPV3-ANO1 interaction positively regulates wound healing in keratinocytes.

doi: 10.1038/s42003-023-04482-1

Figure Lengend Snippet: Fig. 3 TRPV3 and ANO1 interaction in NHEKs. a, b Representative traces of camphor-induced currents in NHEKs using an NMDG-Cl bath solution containing 148 mM chloride (a) or an NMDG-aspartate bath solution containing 4 mM chloride (b). The pipette solution contained 140 mM NMDG-Cl and 100 nM free calcium. The holding potential was −60 mV and ramp-pulses were applied from −100 to +100 mV for 300 ms duration every 5 s. c Comparison of current–voltage relationships of the currents at arrowheads in (a, b). d Calcium imaging of NHEKs upon camphor application with and without (Ca2+ (−)) extracellular calcium. e Comparison of camphor-induced peak currents at −60 mV in an NMDG-Cl bath solution with 2 mM (Ca2+

Article Snippet: The following antibodies were used: rabbit anti-ANO1 antibody (Abcam, ab53213, 1:5 for Western blotting), (Abcam, ab53212, 1:100 for immunoprecipitation), rabbit anti-phospho-ERK (extracellular signal-related kinase) antibody (Cell Signaling Technology, #4370, 1:1000), rabbit anti-phosphop38 antibody (Cell Signaling Technology, #4511, 1:1000), rabbit anti-phospho-JNK (c-Jun N-terminal Kinase) antibody (Cell Signaling Technology, #4668, 1:1000), rabbit anti-ERK antibody (Cell Signaling Technology, #4695, 1:1000), rabbit antip38 antibody (Cell Signaling Technology, #8690, 1:1000), rabbit anti-JNK antibody (Cell Signaling Technology, #9252, 1:1000), mouse anti-β-actin antibody (Abcam, ab6276, 1:2500), rabbit anti-TRPV3 antibody (Cell Signaling Technology, #3484, 1:1000 for Western blotting; 1:50 for immunoprecipitation), anti-rabbit-HRP antibody (Cell Signaling Technology, #7074, 1:2000) and anti-mouse-HRP antibody (Cell Signaling Technology, #7076, 1:2000).

Techniques: Transferring, Comparison, Imaging

Journal: Communications biology

Article Title: TRPV3-ANO1 interaction positively regulates wound healing in keratinocytes.

doi: 10.1038/s42003-023-04482-1

Figure Lengend Snippet: Fig. 4 TRPV3 and ANO1 interaction in mouse primary skin keratinocytes. a, b Representative traces of 10 mM camphor-induced currents in primary keratinocytes isolated from WT (a) and TRPV3−/−(b) mice. All the recordings were performed with standard NMDG-Cl bath and pipette solutions (with 100 nM free calcium in the pipette). The holding potential was −60 mV and ramp-pulses were applied from −100 to +100 mV for 300 ms duration every 3 s. c Comparison of current–voltage relationships. N equals 5 for WT (black) and 4 for TRPV3−/−(gray) keratinocytes. Error bars indicate SEM. d Comparison of densities of the camphor-induced currents in WT (black) and TRPV3−/−(gray) keratinocytes at +60 and −60 mV (means ± SEM). Statistical significance was determined with a Mann–Whitney test. e A representative trace of 10 mM camphor-induced currents with Ani9 inhibition in a WT keratinocyte. A representative trace of 10 mM camphor-induced currents with Ani9 inhibition in a WT keratinocyte. The holding potential was −60 mV and ramp-pulses were applied from −100 to +100 mV for 300 ms duration every 3 s.

Article Snippet: The following antibodies were used: rabbit anti-ANO1 antibody (Abcam, ab53213, 1:5 for Western blotting), (Abcam, ab53212, 1:100 for immunoprecipitation), rabbit anti-phospho-ERK (extracellular signal-related kinase) antibody (Cell Signaling Technology, #4370, 1:1000), rabbit anti-phosphop38 antibody (Cell Signaling Technology, #4511, 1:1000), rabbit anti-phospho-JNK (c-Jun N-terminal Kinase) antibody (Cell Signaling Technology, #4668, 1:1000), rabbit anti-ERK antibody (Cell Signaling Technology, #4695, 1:1000), rabbit antip38 antibody (Cell Signaling Technology, #8690, 1:1000), rabbit anti-JNK antibody (Cell Signaling Technology, #9252, 1:1000), mouse anti-β-actin antibody (Abcam, ab6276, 1:2500), rabbit anti-TRPV3 antibody (Cell Signaling Technology, #3484, 1:1000 for Western blotting; 1:50 for immunoprecipitation), anti-rabbit-HRP antibody (Cell Signaling Technology, #7074, 1:2000) and anti-mouse-HRP antibody (Cell Signaling Technology, #7076, 1:2000).

Techniques: Isolation, Transferring, Comparison, MANN-WHITNEY, Inhibition

Journal: BMC Women's Health

Article Title: Increased capsaicin receptor TRPV1 in skin nerve fibres and related vanilloid receptors TRPV3 and TRPV4 in keratinocytes in human breast pain

doi: 10.1186/1472-6874-5-2

Figure Lengend Snippet: TRPV3-immunoreactivity in breast skin . TOP PANELS: TRPV3-immunoreactive keratinocytes mostly in basal layer and graded from grade 0/negative (top left) to grade 3/strong staining (bottom right). Scale bar = 100 μm BOTTOM PANEL: Scattergram showing grading assessment and a significant increase (*P < 0.05) of TRPV3 immunoreactivity in patients with pain.

Article Snippet: After a further wash in PBS the tissue sections were incubated overnight with affinity purified antibodies to TRPV1 (polyclonal rabbit anti-TRPV1; GlaxoSmithKline, Harlow, UK; 1/5000; 1/10,000), TRPV3 (polyclonal rabbit anti-TRPV3; GlaxoSmithKline, Harlow, UK; 1/1000), TRPV4 (polyclonal rabbit anti-TRPV4; GlaxoSmithKline, Harlow, UK; 1/250; 1/1000), recombinant human NGF (polyclonal rabbit anti-NGF; Genentech, San Francisco, USA; 1/4000) or marker of large and some small calibre nerve fibres (mixed mouse monoclonal antibodies to neurofilaments 200 kD, 70 kD and 57 kD; DAKO cytomation Cambs., UK, 1/50,000; Novocastra, Newcastle upon Tyne, UK, 1/500).

Techniques: Staining

Journal: eNeuro

Article Title: Localization of TRP Channels in Healthy Oral Mucosa from Human Donors

doi: 10.1523/ENEURO.0328-21.2022

Figure Lengend Snippet: Antibodies used in this study

Article Snippet: Rabbit anti-TRPV3 , Alomone Labs , ACC-033 , ACC033AN02 , 1:100 , AB_2040261.

Techniques:

Journal: eNeuro

Article Title: Localization of TRP Channels in Healthy Oral Mucosa from Human Donors

doi: 10.1523/ENEURO.0328-21.2022

Figure Lengend Snippet: Antigen blocking shows specificity of TRP antibodies. A , TRPV3 immunoreactivity was identified in the foreskin epithelium. Antigen pretreatment blocked TRPV3 immunoreactivity ( N = 3 foreskins, paired t test p = 0.022). B , TRPV4 immunoreactivity was identified in the foreskin epithelium and was diminished with antigen pretreatment ( N = 7 foreskins, Wilcoxon matched pairs signed-rank test p = 0.016). Note, removal of outlier data point results in p = 0.02. C , TRPA1 immunoreactivity was identified primarily in the dermal compartment of foreskin tissue and was completely ablated with antigen pretreatment ( N = 4 foreskins or tongues, paired t test, p = 0.0016). D , TRPM8 immunoreactivity was localized primarily to the foreskin epithelium and portions of the dermal compartment. Antigen blocking greatly diminished immunoreactivity ( N = 3 foreskins, paired t test, N = 0.04). E , TRPV1 immunoreactivity was found primarily in intraepithelial nerve fibers of the foreskin. Antigen pretreatment completely abolished TRPV1 nerve fiber immunoreactivity but not the diffuse TRPV1 immunoreactivity of epithelial cells ( N = 3 foreskins, paired t test, N = 0.0004). All graphs show mean ± standard deviation.

Article Snippet: Rabbit anti-TRPV3 , Alomone Labs , ACC-033 , ACC033AN02 , 1:100 , AB_2040261.

Techniques: Blocking Assay, Standard Deviation

Journal: eNeuro

Article Title: Localization of TRP Channels in Healthy Oral Mucosa from Human Donors

doi: 10.1523/ENEURO.0328-21.2022

Figure Lengend Snippet: TRPV3 is expressed in basal layers of oral epithelium. Left column, Tuj1 anti-βIII tubulin (all afferent neurons). Second column, Anti-NFH antibody (myelinated neurons). Third column, Anti-TRPV3 antibody. Right column, Merge with TRP immunoreactivity in yellow, βIII immunoreactivity in cyan, NFH immunoreactivity in magenta. Dashed line indicates epithelia-lamina propria border. A , TRPV3 expression was found in basal epithelial layers of the hard palate as well as in the lamina propria. White arrows denote areas where Merkel cells typically concentrate . Note that Merkel cell afferents are visible with βIII tubulin staining in cyan. B , TRPV3 immunoreactivity was dispersed throughout fungiform epithelium. C , TRPV3 expression was found in basal epithelium of filiform papilla. A TRPV3 negative end bulb of Krause (arrow) is found within the lamina propria of the filiform papilla visualized with expression of βIII tubulin and NFH.

Article Snippet: Rabbit anti-TRPV3 , Alomone Labs , ACC-033 , ACC033AN02 , 1:100 , AB_2040261.

Techniques: Expressing, Staining