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    Structured Review

    Alomone Labs nds
    Nds, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nds/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nds - by Bioz Stars, 2022-01
    94/100 stars

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  • 94
    Alomone Labs anti rat trpv1
    Immunofluorescence for CD88, interleukin-8 RA (IL-8RA) and transient receptor potential cation channel subfamily V member 1 <t>(TRPV1)</t> on 3rd day after administration of doxorubicin into mouse dorsal skin followed by 3-hrs compress. Box plot explanation: The median for each group is shown as a box plot with whiskers from minimum to maximum in (B, D and F). The interquartile range is shown as a box with the median marked as a horizontal line, minimum and maximum from lower and upper quartile represent error bar. (A B) Immunofluorescence for CD88 (A) and IL-8RA (C), and the number (per mm 2 ) of CD88 (B) and IL-8RA (D)-positive cells after 3-hrs cold or hot compresses. Immunofluorescence of TRPV1-positive cells (E) and the number (per mm 2 ) of TRPV1-positive nerve fascicles (F) after 3-hrs cold or hot compresses. [(a) TRPV1 (fluorescein isothiocyanate; FITC), (d) α-smooth muscle actin (α-SMA) (tetramethylrhodamine; TRITC), (g) merge TRPV1 (arrowheads); (b) TRPV1 (FITC), (e) α-SMA (TRITC), (h) merge TRPV1 (arrowheads); (c) TRPV1 (FITC), (f) α-SMA (TRITC), (i) merge TRPV1 (arrowheads)]. (a) control group. (b) cold compress group. (c) hot compress group in (A, C and E). Bar = 50 μm, *: P
    Anti Rat Trpv1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rat trpv1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti rat trpv1 - by Bioz Stars, 2022-01
    94/100 stars
      Buy from Supplier

    99
    Alomone Labs rabbit anti trpv1
    Antagonism of Sig-1R leads to changes in <t>TRPV1</t> expression in HEK293 cells. ( A and B , Left ) Immunodetection of TRPV1 and GAPDH total proteins from HEK293 cells expressing TRPV1 treated for 24 h with 25 µM BD1063 (BD) ( A ) or P4 ( B ) and control (Ctr) [water in A ; methanol (0.25%) in B ]. ( A , Right ) Mean values of TRPV1 for the BD group were 55 ± 6% ( n = 14) with respect to control. *** P
    Rabbit Anti Trpv1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti trpv1/product/Alomone Labs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti trpv1 - by Bioz Stars, 2022-01
    99/100 stars
      Buy from Supplier

    94
    Alomone Labs trpv1 antibody
    <t>TRPV1</t> gene and protein expression using RNA-sequencing data and Western blotting. (A) TRPV1 gene expression in different databases. *P
    Trpv1 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trpv1 antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trpv1 antibody - by Bioz Stars, 2022-01
    94/100 stars
      Buy from Supplier

    Image Search Results


    Immunofluorescence for CD88, interleukin-8 RA (IL-8RA) and transient receptor potential cation channel subfamily V member 1 (TRPV1) on 3rd day after administration of doxorubicin into mouse dorsal skin followed by 3-hrs compress. Box plot explanation: The median for each group is shown as a box plot with whiskers from minimum to maximum in (B, D and F). The interquartile range is shown as a box with the median marked as a horizontal line, minimum and maximum from lower and upper quartile represent error bar. (A B) Immunofluorescence for CD88 (A) and IL-8RA (C), and the number (per mm 2 ) of CD88 (B) and IL-8RA (D)-positive cells after 3-hrs cold or hot compresses. Immunofluorescence of TRPV1-positive cells (E) and the number (per mm 2 ) of TRPV1-positive nerve fascicles (F) after 3-hrs cold or hot compresses. [(a) TRPV1 (fluorescein isothiocyanate; FITC), (d) α-smooth muscle actin (α-SMA) (tetramethylrhodamine; TRITC), (g) merge TRPV1 (arrowheads); (b) TRPV1 (FITC), (e) α-SMA (TRITC), (h) merge TRPV1 (arrowheads); (c) TRPV1 (FITC), (f) α-SMA (TRITC), (i) merge TRPV1 (arrowheads)]. (a) control group. (b) cold compress group. (c) hot compress group in (A, C and E). Bar = 50 μm, *: P

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Effect of cold and hot compress on neutrophilic migration to the site of doxorubicin extravasation

    doi:

    Figure Lengend Snippet: Immunofluorescence for CD88, interleukin-8 RA (IL-8RA) and transient receptor potential cation channel subfamily V member 1 (TRPV1) on 3rd day after administration of doxorubicin into mouse dorsal skin followed by 3-hrs compress. Box plot explanation: The median for each group is shown as a box plot with whiskers from minimum to maximum in (B, D and F). The interquartile range is shown as a box with the median marked as a horizontal line, minimum and maximum from lower and upper quartile represent error bar. (A B) Immunofluorescence for CD88 (A) and IL-8RA (C), and the number (per mm 2 ) of CD88 (B) and IL-8RA (D)-positive cells after 3-hrs cold or hot compresses. Immunofluorescence of TRPV1-positive cells (E) and the number (per mm 2 ) of TRPV1-positive nerve fascicles (F) after 3-hrs cold or hot compresses. [(a) TRPV1 (fluorescein isothiocyanate; FITC), (d) α-smooth muscle actin (α-SMA) (tetramethylrhodamine; TRITC), (g) merge TRPV1 (arrowheads); (b) TRPV1 (FITC), (e) α-SMA (TRITC), (h) merge TRPV1 (arrowheads); (c) TRPV1 (FITC), (f) α-SMA (TRITC), (i) merge TRPV1 (arrowheads)]. (a) control group. (b) cold compress group. (c) hot compress group in (A, C and E). Bar = 50 μm, *: P

    Article Snippet: We then fixed the sections with cold acetone (4°C) for 10 min, followed by addition of 3% skim milk (Morinaga Milk Industry, Tokyo, Japan) dissolved in PBS after washing in 0.01 M phosphate-buffered saline, pH 7.4 (PBS), for 15 min. For double immunofluorescence, slide-mounted tissue sections were incubated with a primary antibody [anti-CD88 (C5aR) antibody (rabbit, polyclonal, Santa Cruz Biotechnology, Santa Cruz, CA), anti-rat IL-8RA antibody (recognizing mouse IL-8RA, goat polyclonal, Santa Cruz Biotechnology), anti-rat TRPV1 (recognizing mouse TRPV1, rabbit, polyclonal, Alomone Labs, Jerusalem, Israel) and mouse monoclonal anti-α-SMA antibody (Clone 1A4, mouse IgG2a, Dako, Glostrup, Denmark)] overnight at 4°C.

    Techniques: Immunofluorescence

    Reverse transcription-polymerase chain reaction (RT-PCR) for CD88, interleukin-8 RA (IL-8RA), and transient receptor potential cation channel subfamily V member 1 (TRPV1). mRNAs of CD88, IL-8RA, and TRPV1 including glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were expressed in skin tissue on the first day after doxorubicin administration. MM: molecular marker, Lane 1; positive control (brain), Lane 2; control (no compress), Lane 3; cold compress, Lane 4; hot compress, Lane 5; negative control.

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Effect of cold and hot compress on neutrophilic migration to the site of doxorubicin extravasation

    doi:

    Figure Lengend Snippet: Reverse transcription-polymerase chain reaction (RT-PCR) for CD88, interleukin-8 RA (IL-8RA), and transient receptor potential cation channel subfamily V member 1 (TRPV1). mRNAs of CD88, IL-8RA, and TRPV1 including glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were expressed in skin tissue on the first day after doxorubicin administration. MM: molecular marker, Lane 1; positive control (brain), Lane 2; control (no compress), Lane 3; cold compress, Lane 4; hot compress, Lane 5; negative control.

    Article Snippet: We then fixed the sections with cold acetone (4°C) for 10 min, followed by addition of 3% skim milk (Morinaga Milk Industry, Tokyo, Japan) dissolved in PBS after washing in 0.01 M phosphate-buffered saline, pH 7.4 (PBS), for 15 min. For double immunofluorescence, slide-mounted tissue sections were incubated with a primary antibody [anti-CD88 (C5aR) antibody (rabbit, polyclonal, Santa Cruz Biotechnology, Santa Cruz, CA), anti-rat IL-8RA antibody (recognizing mouse IL-8RA, goat polyclonal, Santa Cruz Biotechnology), anti-rat TRPV1 (recognizing mouse TRPV1, rabbit, polyclonal, Alomone Labs, Jerusalem, Israel) and mouse monoclonal anti-α-SMA antibody (Clone 1A4, mouse IgG2a, Dako, Glostrup, Denmark)] overnight at 4°C.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Marker, Positive Control, Negative Control

    Recovery of TRPV1 + nerves one week after a sterile epithelial abrasion ( A – L ). The distribution of βIII + TRPV1 + nerves was different across the central ( A – F ) and peripheral cornea ( G – L ). ( M , N ) One-week post-injury, the sum length of TRPV1 + SNT was lower than TRPV1 − SNT in the central cornea ( P

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Distribution of Corneal TRPV1 and Its Association With Immune Cells During Homeostasis and Injury

    doi: 10.1167/iovs.62.9.6

    Figure Lengend Snippet: Recovery of TRPV1 + nerves one week after a sterile epithelial abrasion ( A – L ). The distribution of βIII + TRPV1 + nerves was different across the central ( A – F ) and peripheral cornea ( G – L ). ( M , N ) One-week post-injury, the sum length of TRPV1 + SNT was lower than TRPV1 − SNT in the central cornea ( P

    Article Snippet: They were then incubated overnight with rabbit α-TRPV1 antibody (1:400, Alomone).

    Techniques:

    TRPV1 expression in wild type (WT) and TRPV1 −/− mice. Localization of TRPV1 in corneal nerves and its intracellular localization in the stromal macrophages in wild type mouse corneas ( A, B ), and absent in the TRPV1 −/− mouse cornea ( C, D ). The punctate staining in the epithelium was present in the TRPV1 −/− mice, likely representing nonspecific background staining. Scale bars represent 50 µm for all images. ( E, F ) Punctate TRPV1 signals in the CD45 + stromal macrophages were present in the WT but absent in the TRPV1 −/− mouse corneas ( G ).

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Distribution of Corneal TRPV1 and Its Association With Immune Cells During Homeostasis and Injury

    doi: 10.1167/iovs.62.9.6

    Figure Lengend Snippet: TRPV1 expression in wild type (WT) and TRPV1 −/− mice. Localization of TRPV1 in corneal nerves and its intracellular localization in the stromal macrophages in wild type mouse corneas ( A, B ), and absent in the TRPV1 −/− mouse cornea ( C, D ). The punctate staining in the epithelium was present in the TRPV1 −/− mice, likely representing nonspecific background staining. Scale bars represent 50 µm for all images. ( E, F ) Punctate TRPV1 signals in the CD45 + stromal macrophages were present in the WT but absent in the TRPV1 −/− mouse corneas ( G ).

    Article Snippet: They were then incubated overnight with rabbit α-TRPV1 antibody (1:400, Alomone).

    Techniques: Expressing, Mouse Assay, Staining

    TRPV1 localization in nonneuronal cell bodies and their partial overlap with lysosomes of the stromal macrophages. ( A–C ) βIII + stromal nerve trunks expressed TRPV1. Intracellular TRPV1 staining was present in the macrophages that were in various depths of the stroma (color-coded depth projection, anterior: blue/purple; posterior: yellow) ( B , arrowheads; C , asterisks). ( D ) Absence of TRPV1 staining in tissues treated with peptide control antigen alongside positive staining of stromal CD45 + cells (red). ( E1–4 ) High resolution image of TRPV1 staining within the CD45 + stromal macrophages. ( F1–4 ) TRPV1 partially overlapped with the macrophage intracellular compartment that expresses CD68. ( G ) Orthogonal view of a CD45 + TRPV1 + stromal macrophage displaying the nonneuronal TRPV1 localization in both XY and XZ planes of the z-stack. ( H ) Orthogonal view of a CD68 + TRPV1 + stromal macrophage shows the overlap between lysosomes and TRPV1 proteins. Scale bars represent 100 µm ( A – D ) and 10 µm (E1 – 4, F1 – 4, G, H).

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Distribution of Corneal TRPV1 and Its Association With Immune Cells During Homeostasis and Injury

    doi: 10.1167/iovs.62.9.6

    Figure Lengend Snippet: TRPV1 localization in nonneuronal cell bodies and their partial overlap with lysosomes of the stromal macrophages. ( A–C ) βIII + stromal nerve trunks expressed TRPV1. Intracellular TRPV1 staining was present in the macrophages that were in various depths of the stroma (color-coded depth projection, anterior: blue/purple; posterior: yellow) ( B , arrowheads; C , asterisks). ( D ) Absence of TRPV1 staining in tissues treated with peptide control antigen alongside positive staining of stromal CD45 + cells (red). ( E1–4 ) High resolution image of TRPV1 staining within the CD45 + stromal macrophages. ( F1–4 ) TRPV1 partially overlapped with the macrophage intracellular compartment that expresses CD68. ( G ) Orthogonal view of a CD45 + TRPV1 + stromal macrophage displaying the nonneuronal TRPV1 localization in both XY and XZ planes of the z-stack. ( H ) Orthogonal view of a CD68 + TRPV1 + stromal macrophage shows the overlap between lysosomes and TRPV1 proteins. Scale bars represent 100 µm ( A – D ) and 10 µm (E1 – 4, F1 – 4, G, H).

    Article Snippet: They were then incubated overnight with rabbit α-TRPV1 antibody (1:400, Alomone).

    Techniques: Staining

    Epithelial TRPV1 + nerves and resident dendritic cells (DCs) under steady state conditions. ( A ) Representative images from the peripheral corneal epithelium showing resident intraepithelial CD45 + DCs in close proximity to TRPV1 + axons (asterisk), while some DCs did not appear to associate with TRPV1 + axons (arrowheads). ( B ) Orthogonal view of an intraepithelial CD45 + DC highlights the contacts between the TRPV1 + axons and DC in XYZ planes. ( C, D ) The absence of TRPV1 nerve staining in the peptide control validates the positive corneal nerve staining for TRPV1. ( E–H ) The resident DCs were immunopositive for CD11c and MHC class II. Scale bars represent 50 µm ( A , C , D ) and 10 µm ( E , F , G , H ).

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Distribution of Corneal TRPV1 and Its Association With Immune Cells During Homeostasis and Injury

    doi: 10.1167/iovs.62.9.6

    Figure Lengend Snippet: Epithelial TRPV1 + nerves and resident dendritic cells (DCs) under steady state conditions. ( A ) Representative images from the peripheral corneal epithelium showing resident intraepithelial CD45 + DCs in close proximity to TRPV1 + axons (asterisk), while some DCs did not appear to associate with TRPV1 + axons (arrowheads). ( B ) Orthogonal view of an intraepithelial CD45 + DC highlights the contacts between the TRPV1 + axons and DC in XYZ planes. ( C, D ) The absence of TRPV1 nerve staining in the peptide control validates the positive corneal nerve staining for TRPV1. ( E–H ) The resident DCs were immunopositive for CD11c and MHC class II. Scale bars represent 50 µm ( A , C , D ) and 10 µm ( E , F , G , H ).

    Article Snippet: They were then incubated overnight with rabbit α-TRPV1 antibody (1:400, Alomone).

    Techniques: Staining

    Intraepithelial CD45 + dendriform cells in intact and injured corneas. ( A–D ) βIII, TRPV1, and CD45 staining demonstrate TRPV1 + and TRPV1 − nerve populations and their anatomical associations with DCs in the central and peripheral epithelium of intact ( A , B ) and injured corneas ( C , D ). ( E–L ) 3D image reconstructions show DC-nerve contacts between epithelial CD45 + DCs and βIII + TRPV1 + SBNP nerve axons (green: βIII; white: TRPV1). ( G–H ) DC-nerve complexes between DC and nerve axons (white area: βIII; yellow: TRPV1) were found within the DC dendrite. ( I–L ) One week after the epithelial abrasion, the degree of neuroimmune contacts between the DCs and nerve axons was similar to that observed in intact corneas (white area: βIII; blue: TRPV1) and DC dendrites. ( M, N ) The percentage of TRPV1 + nerve-associated DC was significantly higher across both central and peripheral corneas in the injured group compared to the intact control corneas ( P

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Distribution of Corneal TRPV1 and Its Association With Immune Cells During Homeostasis and Injury

    doi: 10.1167/iovs.62.9.6

    Figure Lengend Snippet: Intraepithelial CD45 + dendriform cells in intact and injured corneas. ( A–D ) βIII, TRPV1, and CD45 staining demonstrate TRPV1 + and TRPV1 − nerve populations and their anatomical associations with DCs in the central and peripheral epithelium of intact ( A , B ) and injured corneas ( C , D ). ( E–L ) 3D image reconstructions show DC-nerve contacts between epithelial CD45 + DCs and βIII + TRPV1 + SBNP nerve axons (green: βIII; white: TRPV1). ( G–H ) DC-nerve complexes between DC and nerve axons (white area: βIII; yellow: TRPV1) were found within the DC dendrite. ( I–L ) One week after the epithelial abrasion, the degree of neuroimmune contacts between the DCs and nerve axons was similar to that observed in intact corneas (white area: βIII; blue: TRPV1) and DC dendrites. ( M, N ) The percentage of TRPV1 + nerve-associated DC was significantly higher across both central and peripheral corneas in the injured group compared to the intact control corneas ( P

    Article Snippet: They were then incubated overnight with rabbit α-TRPV1 antibody (1:400, Alomone).

    Techniques: Staining

    Topographical distribution of TRPV1 + nerves in the intact mouse cornea. ( A–L ) Distribution of βIII + ( A , D , G , and J ) and TRPV1 + ( B , E , H , and K ) superficial nerve terminals (SNT) and subbasal nerve plexus (SBNP) in the central ( A–F ) and peripheral corneal epithelium ( G – L ). False colored merged images demonstrate the relationship between βIII + TRPV1 + (cyan) and βIII + TRPV1 − (magenta) nerve fibers ( C , F , I , and L ). ( M – P ) The sum length of TRPV1 + SNT was lower than TRPV1 − SNT in the central cornea ( P

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Distribution of Corneal TRPV1 and Its Association With Immune Cells During Homeostasis and Injury

    doi: 10.1167/iovs.62.9.6

    Figure Lengend Snippet: Topographical distribution of TRPV1 + nerves in the intact mouse cornea. ( A–L ) Distribution of βIII + ( A , D , G , and J ) and TRPV1 + ( B , E , H , and K ) superficial nerve terminals (SNT) and subbasal nerve plexus (SBNP) in the central ( A–F ) and peripheral corneal epithelium ( G – L ). False colored merged images demonstrate the relationship between βIII + TRPV1 + (cyan) and βIII + TRPV1 − (magenta) nerve fibers ( C , F , I , and L ). ( M – P ) The sum length of TRPV1 + SNT was lower than TRPV1 − SNT in the central cornea ( P

    Article Snippet: They were then incubated overnight with rabbit α-TRPV1 antibody (1:400, Alomone).

    Techniques:

    Stromal TRPV1 + nerves and CD45 + TRPV1 + macrophages in intact and injured corneas. ( A–D ) Superimposed images of βIII, TRPV1, and CD45 staining demonstrate TRPV1 + and TRPV1 − nerves and their anatomical interaction with macrophages in central and peripheral stroma of intact ( A , B ) and injured corneas ( C , D ). ( E–H ) The sum lengths of TRPV1 + stromal nerves were higher in both central and peripheral regions of the intact corneas ( P

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Distribution of Corneal TRPV1 and Its Association With Immune Cells During Homeostasis and Injury

    doi: 10.1167/iovs.62.9.6

    Figure Lengend Snippet: Stromal TRPV1 + nerves and CD45 + TRPV1 + macrophages in intact and injured corneas. ( A–D ) Superimposed images of βIII, TRPV1, and CD45 staining demonstrate TRPV1 + and TRPV1 − nerves and their anatomical interaction with macrophages in central and peripheral stroma of intact ( A , B ) and injured corneas ( C , D ). ( E–H ) The sum lengths of TRPV1 + stromal nerves were higher in both central and peripheral regions of the intact corneas ( P

    Article Snippet: They were then incubated overnight with rabbit α-TRPV1 antibody (1:400, Alomone).

    Techniques: Staining

    Antagonism of Sig-1R leads to changes in TRPV1 expression in HEK293 cells. ( A and B , Left ) Immunodetection of TRPV1 and GAPDH total proteins from HEK293 cells expressing TRPV1 treated for 24 h with 25 µM BD1063 (BD) ( A ) or P4 ( B ) and control (Ctr) [water in A ; methanol (0.25%) in B ]. ( A , Right ) Mean values of TRPV1 for the BD group were 55 ± 6% ( n = 14) with respect to control. *** P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: TRPV1 channels and the progesterone receptor Sig-1R interact to regulate pain

    doi: 10.1073/pnas.1715972115

    Figure Lengend Snippet: Antagonism of Sig-1R leads to changes in TRPV1 expression in HEK293 cells. ( A and B , Left ) Immunodetection of TRPV1 and GAPDH total proteins from HEK293 cells expressing TRPV1 treated for 24 h with 25 µM BD1063 (BD) ( A ) or P4 ( B ) and control (Ctr) [water in A ; methanol (0.25%) in B ]. ( A , Right ) Mean values of TRPV1 for the BD group were 55 ± 6% ( n = 14) with respect to control. *** P

    Article Snippet: Membranes were blocked with 6% nonfat dry milk in PBS and Tween-20 0.1% (PBS-T) and incubated overnight with goat polyclonal anti-TRPV1 (P-19, sc12498; Santa Cruz Biotechnology) or rabbit anti-TRPV1 (ACC-030 and ACC-029; Alomone Laboratories), diluted 1/1,000 in PBS-T with 3% nonfat dry milk.

    Techniques: Expressing, Immunodetection

    TRPV1-mediated pain is dependent upon Sig-1R activity. ( A ) Response to 2.8-µg capsaicin injection into to the hind paw of male mice that had been pretreated for 24 h by intrascapularly injecting either vehicle (water) or a Sig-1R antagonist (BD1063) (32 mg/kg in water). PLT in response to capsaicin were 50 ± 5 s for vehicle-pretreated ( n = 6) and 24 ± 4 s for BD1063-pretreated ( n = 6) mice; *** P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: TRPV1 channels and the progesterone receptor Sig-1R interact to regulate pain

    doi: 10.1073/pnas.1715972115

    Figure Lengend Snippet: TRPV1-mediated pain is dependent upon Sig-1R activity. ( A ) Response to 2.8-µg capsaicin injection into to the hind paw of male mice that had been pretreated for 24 h by intrascapularly injecting either vehicle (water) or a Sig-1R antagonist (BD1063) (32 mg/kg in water). PLT in response to capsaicin were 50 ± 5 s for vehicle-pretreated ( n = 6) and 24 ± 4 s for BD1063-pretreated ( n = 6) mice; *** P

    Article Snippet: Membranes were blocked with 6% nonfat dry milk in PBS and Tween-20 0.1% (PBS-T) and incubated overnight with goat polyclonal anti-TRPV1 (P-19, sc12498; Santa Cruz Biotechnology) or rabbit anti-TRPV1 (ACC-030 and ACC-029; Alomone Laboratories), diluted 1/1,000 in PBS-T with 3% nonfat dry milk.

    Techniques: Activity Assay, Injection, Mouse Assay

    Colocalization of TRPV1 and Sig-1R in HEK293 cells and DRG neurons by confocal microscopy. ( A ) Panels 1 and 2 show signals detected for the TRPV1-mCherry ( 1 , red) and Sig-1R ( 2 , green) proteins. Panel 3 shows labeling of the ER by rabbit anti-calnexin and anti-rabbit Cy5 antibodies (blue). Colocalization of TRPV1 and Sig-1R in the ER is shown in panel 4 (arrow indicates the merge in white). ( B ) Panels 1 and 2 are the same as in A . Panel 3 shows the plasma membrane labelled with CellMask Deep Red (blue). Panel 4 shows the merged image (purple) and strong localization of TRPV1 (red) to the plasma membrane. ( C ) Colocalization of endogenous TRPV1 and Sig-1R in primary cultures of mouse DRG neurons. TRPV1 immunodetection with goat anti-TRPV1 and secondary anti-goat Alexa Fluor 488 antibodies (green, panel 1 ) and Sig-1R immunodetection with rabbit anti–Sig-1R and secondary anti-rabbit Alexa Fluor 594 antibodies (red, panel 2 ). Merge (yellow, panel 3 ) shows colocalization of both proteins. (Scale bars: 10 μm.) All arrows indicate colocalized signals.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: TRPV1 channels and the progesterone receptor Sig-1R interact to regulate pain

    doi: 10.1073/pnas.1715972115

    Figure Lengend Snippet: Colocalization of TRPV1 and Sig-1R in HEK293 cells and DRG neurons by confocal microscopy. ( A ) Panels 1 and 2 show signals detected for the TRPV1-mCherry ( 1 , red) and Sig-1R ( 2 , green) proteins. Panel 3 shows labeling of the ER by rabbit anti-calnexin and anti-rabbit Cy5 antibodies (blue). Colocalization of TRPV1 and Sig-1R in the ER is shown in panel 4 (arrow indicates the merge in white). ( B ) Panels 1 and 2 are the same as in A . Panel 3 shows the plasma membrane labelled with CellMask Deep Red (blue). Panel 4 shows the merged image (purple) and strong localization of TRPV1 (red) to the plasma membrane. ( C ) Colocalization of endogenous TRPV1 and Sig-1R in primary cultures of mouse DRG neurons. TRPV1 immunodetection with goat anti-TRPV1 and secondary anti-goat Alexa Fluor 488 antibodies (green, panel 1 ) and Sig-1R immunodetection with rabbit anti–Sig-1R and secondary anti-rabbit Alexa Fluor 594 antibodies (red, panel 2 ). Merge (yellow, panel 3 ) shows colocalization of both proteins. (Scale bars: 10 μm.) All arrows indicate colocalized signals.

    Article Snippet: Membranes were blocked with 6% nonfat dry milk in PBS and Tween-20 0.1% (PBS-T) and incubated overnight with goat polyclonal anti-TRPV1 (P-19, sc12498; Santa Cruz Biotechnology) or rabbit anti-TRPV1 (ACC-030 and ACC-029; Alomone Laboratories), diluted 1/1,000 in PBS-T with 3% nonfat dry milk.

    Techniques: Confocal Microscopy, Labeling, Immunodetection

    Antagonism of Sig-1R affects TRPV1 expression and its role in pain. ( A ) Nonpregnant and pregnant (15 d of gestation) mice were intradermally injected into to a hind paw with vehicle- or capsaicin (2.8 µg)-containing solutions. PLTs for vehicle injections of nonpregnant ( n = 18) and pregnant females ( n = 12) were 5.2 ±1.2 s and 2.5 ± 0.9 s, respectively. For capsaicin injection PLTs were 43 ± 2 s in nonpregnant females and 24 ± 1 s in pregnant females. *** P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: TRPV1 channels and the progesterone receptor Sig-1R interact to regulate pain

    doi: 10.1073/pnas.1715972115

    Figure Lengend Snippet: Antagonism of Sig-1R affects TRPV1 expression and its role in pain. ( A ) Nonpregnant and pregnant (15 d of gestation) mice were intradermally injected into to a hind paw with vehicle- or capsaicin (2.8 µg)-containing solutions. PLTs for vehicle injections of nonpregnant ( n = 18) and pregnant females ( n = 12) were 5.2 ±1.2 s and 2.5 ± 0.9 s, respectively. For capsaicin injection PLTs were 43 ± 2 s in nonpregnant females and 24 ± 1 s in pregnant females. *** P

    Article Snippet: Membranes were blocked with 6% nonfat dry milk in PBS and Tween-20 0.1% (PBS-T) and incubated overnight with goat polyclonal anti-TRPV1 (P-19, sc12498; Santa Cruz Biotechnology) or rabbit anti-TRPV1 (ACC-030 and ACC-029; Alomone Laboratories), diluted 1/1,000 in PBS-T with 3% nonfat dry milk.

    Techniques: Expressing, Mouse Assay, Injection

    TRPV1 and Sig-1R form protein–protein complexes. ( A , Left ) Coimmunoprecipitation of TRPV1 with Sig-1R antibody and immunodetection of TRPV1 ( Upper ) and Sig-1R ( Lower ). ( Right ) Coimmunoprecipitation of Sig-1R with TRPV1 antibody and immunodetection for TRPV1 ( Upper ) and Sig-1R ( Lower ). IgG served as negative control. The figure is representative of three independent experiments. Input is 1/10th of the total protein used in each immunoprecipitation. ( B ) Immunoprecipitation was carried out with TRPV1 antibody for the total protein from control cells (Ctr, Left ) and BD1063-treated cells ( Right ). Differences between control and BD1063-treated cells were determined in the signal for immunoprecipitated Sig-1R (lane 5). The figure is representative of three independent experiments. ( C ) Immunoprecipitation was carried out as above with total protein from control cells ( Left ) and P4-treated cells ( Right ). Differences between control and P4-treated cells were determined in the signal for immunoprecipitated Sig-1R (lane 5). The figure is representative of three independent experiments. ( D and E ) FRET between TRPV1-mCherry and Sig-1R-GFP. The ratio A o is the fractional excitation of mCherry in the absence of donor (GFP), and its average value is indicated by the dashed line. A value of ratio A, when both donor and acceptor are present, greater than ratio A o is indicative of FRET. The coexpression of TRPV1 and Sig-1R produces an apparent FRET efficiency of 0.49 ± 0.02 ( n = 17). ( D ) Incubation of the cells in the presence of 25 µM BD1063 reduces the apparent FRET efficiency to 0.42 ± 0.02 ( n = 6), which is not different from the ratio A o . ( E ) Treatment with 25 µM progesterone (P4) has an effect similar to BD1063. Control apparent FRET efficiency is 0.57 ± 0.03 ( n = 10), and progesterone treatment reduces it to 0.45 ± 0.02 ( n = 10). ( F ) Western blot analysis for deletion constructs of TRPV1 protein shows the bands corresponding to the expected sizes: TRPV1-ΔNH2 (58 kDa, Left ), TRPV1-ΔCOOH (77 kDa, Center ), and TRPV1-TM1-6 (28 kDa, Right ). ( G ) Coimmunoprecipitation of different TRPV1 deletion constructs with Sig-1R. Coimmunoprecipitation of the TRPV1-ΔNH 2 ( Top ) and TRPV1-ΔCOOH ( Middle ) deletion constructs and of TM1-6 transmembrane domains only ( Bottom ) with Sig-1R. Each deletion construct of TRPV1 was coimmunoprecipitated with Sig-1R-GFP (IP) by using a specific antibody against a specific epitope of the segment.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: TRPV1 channels and the progesterone receptor Sig-1R interact to regulate pain

    doi: 10.1073/pnas.1715972115

    Figure Lengend Snippet: TRPV1 and Sig-1R form protein–protein complexes. ( A , Left ) Coimmunoprecipitation of TRPV1 with Sig-1R antibody and immunodetection of TRPV1 ( Upper ) and Sig-1R ( Lower ). ( Right ) Coimmunoprecipitation of Sig-1R with TRPV1 antibody and immunodetection for TRPV1 ( Upper ) and Sig-1R ( Lower ). IgG served as negative control. The figure is representative of three independent experiments. Input is 1/10th of the total protein used in each immunoprecipitation. ( B ) Immunoprecipitation was carried out with TRPV1 antibody for the total protein from control cells (Ctr, Left ) and BD1063-treated cells ( Right ). Differences between control and BD1063-treated cells were determined in the signal for immunoprecipitated Sig-1R (lane 5). The figure is representative of three independent experiments. ( C ) Immunoprecipitation was carried out as above with total protein from control cells ( Left ) and P4-treated cells ( Right ). Differences between control and P4-treated cells were determined in the signal for immunoprecipitated Sig-1R (lane 5). The figure is representative of three independent experiments. ( D and E ) FRET between TRPV1-mCherry and Sig-1R-GFP. The ratio A o is the fractional excitation of mCherry in the absence of donor (GFP), and its average value is indicated by the dashed line. A value of ratio A, when both donor and acceptor are present, greater than ratio A o is indicative of FRET. The coexpression of TRPV1 and Sig-1R produces an apparent FRET efficiency of 0.49 ± 0.02 ( n = 17). ( D ) Incubation of the cells in the presence of 25 µM BD1063 reduces the apparent FRET efficiency to 0.42 ± 0.02 ( n = 6), which is not different from the ratio A o . ( E ) Treatment with 25 µM progesterone (P4) has an effect similar to BD1063. Control apparent FRET efficiency is 0.57 ± 0.03 ( n = 10), and progesterone treatment reduces it to 0.45 ± 0.02 ( n = 10). ( F ) Western blot analysis for deletion constructs of TRPV1 protein shows the bands corresponding to the expected sizes: TRPV1-ΔNH2 (58 kDa, Left ), TRPV1-ΔCOOH (77 kDa, Center ), and TRPV1-TM1-6 (28 kDa, Right ). ( G ) Coimmunoprecipitation of different TRPV1 deletion constructs with Sig-1R. Coimmunoprecipitation of the TRPV1-ΔNH 2 ( Top ) and TRPV1-ΔCOOH ( Middle ) deletion constructs and of TM1-6 transmembrane domains only ( Bottom ) with Sig-1R. Each deletion construct of TRPV1 was coimmunoprecipitated with Sig-1R-GFP (IP) by using a specific antibody against a specific epitope of the segment.

    Article Snippet: Membranes were blocked with 6% nonfat dry milk in PBS and Tween-20 0.1% (PBS-T) and incubated overnight with goat polyclonal anti-TRPV1 (P-19, sc12498; Santa Cruz Biotechnology) or rabbit anti-TRPV1 (ACC-030 and ACC-029; Alomone Laboratories), diluted 1/1,000 in PBS-T with 3% nonfat dry milk.

    Techniques: Immunodetection, Negative Control, Immunoprecipitation, Incubation, Western Blot, Construct

    TRPV1 gene and protein expression using RNA-sequencing data and Western blotting. (A) TRPV1 gene expression in different databases. *P

    Journal: medRxiv

    Article Title: Optimized Flow Cytometric Detection of Transient Receptor Potential Vanilloid-1 (TRPV1) in Human Hematological Malignancies

    doi: 10.1101/2021.08.04.21261521

    Figure Lengend Snippet: TRPV1 gene and protein expression using RNA-sequencing data and Western blotting. (A) TRPV1 gene expression in different databases. *P

    Article Snippet: Exclusion of TRPV1 antibody was used as an additional negative control.

    Techniques: Expressing, RNA Sequencing Assay, Western Blot

    TRPV1 expression using Flow Cytometry. (A) Overview structure of TRPV1 and the 3 antibodies binding sites. Assessment of anti-TRPV1 antibodies: (B) SC-20813, (C) ACC-030, and (D) LS-C150735. Panel D demonstrates clear separation of isotype control from TRPV1 signals in THP-1, U266B1, U937 cells and healthy human lymphocytes using LS-C150735. (E) TRPV1 expression in all patients with hematological malignancies and healthy controls (n=20). TRPV1 in MM, B-NHL and other hematological malignancies group (treated and de novo ) were compared to control. Median fluorescence intensity (MFI) value for individual patients expressed using violin plots with the mean and standard deviation also shown. Patients samples were also expressed as a ratio (MFI patient: MFI control). Overall TRPV1 was significantly higher in all patients with hematological malignancies compared to healthy control (P=0.0351, unpaired t-test) with B-NHL patients having significantly higher TRPV1 compared to control (P= 0.0294, Dunnet’s test)

    Journal: medRxiv

    Article Title: Optimized Flow Cytometric Detection of Transient Receptor Potential Vanilloid-1 (TRPV1) in Human Hematological Malignancies

    doi: 10.1101/2021.08.04.21261521

    Figure Lengend Snippet: TRPV1 expression using Flow Cytometry. (A) Overview structure of TRPV1 and the 3 antibodies binding sites. Assessment of anti-TRPV1 antibodies: (B) SC-20813, (C) ACC-030, and (D) LS-C150735. Panel D demonstrates clear separation of isotype control from TRPV1 signals in THP-1, U266B1, U937 cells and healthy human lymphocytes using LS-C150735. (E) TRPV1 expression in all patients with hematological malignancies and healthy controls (n=20). TRPV1 in MM, B-NHL and other hematological malignancies group (treated and de novo ) were compared to control. Median fluorescence intensity (MFI) value for individual patients expressed using violin plots with the mean and standard deviation also shown. Patients samples were also expressed as a ratio (MFI patient: MFI control). Overall TRPV1 was significantly higher in all patients with hematological malignancies compared to healthy control (P=0.0351, unpaired t-test) with B-NHL patients having significantly higher TRPV1 compared to control (P= 0.0294, Dunnet’s test)

    Article Snippet: Exclusion of TRPV1 antibody was used as an additional negative control.

    Techniques: Expressing, Flow Cytometry, Binding Assay, Fluorescence, Standard Deviation