polyclonal anti mas receptor antibody  (Alomone Labs)


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    Structured Review

    Alomone Labs polyclonal anti mas receptor antibody
    Polyclonal Anti Mas Receptor Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polyclonal anti mas receptor antibody/product/Alomone Labs
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    polyclonal anti mas receptor antibody - by Bioz Stars, 2022-08
    85/100 stars

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    Alomone Labs anti mrgprd gpcr tgr7 antibody
    The expression and functional activity of pain- and itch-related receptor <t>MrgprD</t> are upregulated in DRG neurons from oxazolone-induced ACD model mice. A Heatmap showing the expression of Mrgpr family genes identified in DRG of oxazolone versus control group mice. n = 4 mice per group. B qPCR validations of the expression of three itch- or pain-related Mrgpr genes, namely, Mrgprx1 , Mrgpra3 , and Mrgprd . n = 6–8 mice per group. * p
    Anti Mrgprd Gpcr Tgr7 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/anti mrgprd gpcr tgr7 antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti mrgprd gpcr tgr7 antibody - by Bioz Stars, 2022-08
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    94
    Alomone Labs anti trpc6
    Western blot ( WB ) of HEK 293T cells transfected ( Tx'd ) with Epo-R, BFP-TRPC3, and <t>BFP-TRPC6.</t> HEK 293T cells were cotransfected with Epo-R, and BFP-TRPC3, BFP-TRPC6, or both. Equivalent amounts of protein lysates were loaded in each lane, and Western blotting was performed with anti-TRC3, anti-TRPC6, or anti-Epo-R antibodies, followed by ECL.
    Anti Trpc6, 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 trpc6/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    93
    Alomone Labs rabbit polyclonal anti angiotensin 1 7 mas receptor
    The correlation between the plasma level of the total alternative renin angiotensin system activity (Alt-S) and the weight of the visceral fat ( a ) and body weight ( b ). The correlation between the plasma level of angiotensin 1–7 (Ang 1–7) and the weight of the visceral fat ( c ) and body weight ( d ) in SHRs and SHRs treated with MLN-4760 (SHR + MLN).
    Rabbit Polyclonal Anti Angiotensin 1 7 Mas Receptor, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti angiotensin 1 7 mas receptor/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
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    The expression and functional activity of pain- and itch-related receptor MrgprD are upregulated in DRG neurons from oxazolone-induced ACD model mice. A Heatmap showing the expression of Mrgpr family genes identified in DRG of oxazolone versus control group mice. n = 4 mice per group. B qPCR validations of the expression of three itch- or pain-related Mrgpr genes, namely, Mrgprx1 , Mrgpra3 , and Mrgprd . n = 6–8 mice per group. * p

    Journal: Cellular & Molecular Biology Letters

    Article Title: Exploring neuronal mechanisms involved in the scratching behavior of a mouse model of allergic contact dermatitis by transcriptomics

    doi: 10.1186/s11658-022-00316-w

    Figure Lengend Snippet: The expression and functional activity of pain- and itch-related receptor MrgprD are upregulated in DRG neurons from oxazolone-induced ACD model mice. A Heatmap showing the expression of Mrgpr family genes identified in DRG of oxazolone versus control group mice. n = 4 mice per group. B qPCR validations of the expression of three itch- or pain-related Mrgpr genes, namely, Mrgprx1 , Mrgpra3 , and Mrgprd . n = 6–8 mice per group. * p

    Article Snippet: We used the following antibodies: rabbit anti-CD3 (1:200, no. MA1-90582, Thermo Fisher, USA), rabbit anti-MrgprD (1:200, no. AMR-061, Alomone, Israel), and mouse anti-NeuN (1:300, no. ab104224, Abcam, UK).

    Techniques: Expressing, Functional Assay, Activity Assay, Mouse Assay, Real-time Polymerase Chain Reaction

    Western blot ( WB ) of HEK 293T cells transfected ( Tx'd ) with Epo-R, BFP-TRPC3, and BFP-TRPC6. HEK 293T cells were cotransfected with Epo-R, and BFP-TRPC3, BFP-TRPC6, or both. Equivalent amounts of protein lysates were loaded in each lane, and Western blotting was performed with anti-TRC3, anti-TRPC6, or anti-Epo-R antibodies, followed by ECL.

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Activation by Erythropoietin Is Modulated by TRPC6

    doi: 10.1074/jbc.M804734200

    Figure Lengend Snippet: Western blot ( WB ) of HEK 293T cells transfected ( Tx'd ) with Epo-R, BFP-TRPC3, and BFP-TRPC6. HEK 293T cells were cotransfected with Epo-R, and BFP-TRPC3, BFP-TRPC6, or both. Equivalent amounts of protein lysates were loaded in each lane, and Western blotting was performed with anti-TRC3, anti-TRPC6, or anti-Epo-R antibodies, followed by ECL.

    Article Snippet: Blots were incubated with anti-TRPC3 (1:400, Alomone Labs, Jerusalem, Israel), anti-TRPC3 (C) targeted to the human C-terminal sequence RRRRLQDIEMGMGNSKSRLN (1:1000, Bethyl Laboratories, Inc., Montgomery, TX) , anti-TRPC3 (N) targeted to the murine N-terminal sequence LNGDLESAEPLERHGHKASL (1:1000, Bethyl Laboratories, Inc.) , anti-TRPC6 (1:250, Alomone Labs; 1:500, Abcam, Inc., Cambridge, MA), anti-V5-HRP (1:10,000, Invitrogen), anti-FLAG (1:1000, Sigma), anti-PLCγ (1:1000, SC-81, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), anti-Epo-R (1:1000, SC697, Santa Cruz Biotechnology, Inc.), anti-actin (1:10,000, Sigma), anti-tubulin (1:10,000, Sigma) antibodies, or streptavidin-HRP (Pierce).

    Techniques: Western Blot, Transfection

    Interaction of transfected and endogenous TRPC3 and TRPC6. A, V5-TRPC3 and/or FLAG-TRPC6 were expressed in HEK 293T cells with Epo-R. Immunoprecipitation ( IP ) was performed on lysates with anti-V5 antibody or anti-FLAG-agarose. Western blotting ( WB ) was performed after immunoprecipitation with anti-V5 or anti-FLAG antibodies. Representative results of five experiments are shown. B, immunoprecipitation was performed on lysates from TF-1 erythroid cells with anti-TRPC3 or anti-TRPC6 antibody to detect endogenous interactions. Western blotting was performed after immunoprecipitation with anti-TRPC3 or anti-TRPC3 antibodies. Representative results of two experiments are shown.

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Activation by Erythropoietin Is Modulated by TRPC6

    doi: 10.1074/jbc.M804734200

    Figure Lengend Snippet: Interaction of transfected and endogenous TRPC3 and TRPC6. A, V5-TRPC3 and/or FLAG-TRPC6 were expressed in HEK 293T cells with Epo-R. Immunoprecipitation ( IP ) was performed on lysates with anti-V5 antibody or anti-FLAG-agarose. Western blotting ( WB ) was performed after immunoprecipitation with anti-V5 or anti-FLAG antibodies. Representative results of five experiments are shown. B, immunoprecipitation was performed on lysates from TF-1 erythroid cells with anti-TRPC3 or anti-TRPC6 antibody to detect endogenous interactions. Western blotting was performed after immunoprecipitation with anti-TRPC3 or anti-TRPC3 antibodies. Representative results of two experiments are shown.

    Article Snippet: Blots were incubated with anti-TRPC3 (1:400, Alomone Labs, Jerusalem, Israel), anti-TRPC3 (C) targeted to the human C-terminal sequence RRRRLQDIEMGMGNSKSRLN (1:1000, Bethyl Laboratories, Inc., Montgomery, TX) , anti-TRPC3 (N) targeted to the murine N-terminal sequence LNGDLESAEPLERHGHKASL (1:1000, Bethyl Laboratories, Inc.) , anti-TRPC6 (1:250, Alomone Labs; 1:500, Abcam, Inc., Cambridge, MA), anti-V5-HRP (1:10,000, Invitrogen), anti-FLAG (1:1000, Sigma), anti-PLCγ (1:1000, SC-81, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), anti-Epo-R (1:1000, SC697, Santa Cruz Biotechnology, Inc.), anti-actin (1:10,000, Sigma), anti-tubulin (1:10,000, Sigma) antibodies, or streptavidin-HRP (Pierce).

    Techniques: Transfection, Immunoprecipitation, Western Blot

    Schema of TRPC3/TRPC6 chimeras. A, schematic model of TRPC3-C6C, TRPC6-C3N, and TRPC6-C3C chimeras. B, Western blot ( WB ) of lysates from HEK 293T cells transfected with BFP-TRPC3, BFP-TRPC6, BFP-TRPC6-C3C, BFP-TRPC6-C3N, or BFP-TRPC3-C6C. Blots were probed with antibodies that recognize the C terminus of human TRPC3 (anti-TRPC3 (C)), the N terminus of murine and human TRPC3 (anti-TRPC3 (N)), or the N terminus of TRPC6 (anti-TRPC6 (N)). Representative results of two experiments are shown.

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Activation by Erythropoietin Is Modulated by TRPC6

    doi: 10.1074/jbc.M804734200

    Figure Lengend Snippet: Schema of TRPC3/TRPC6 chimeras. A, schematic model of TRPC3-C6C, TRPC6-C3N, and TRPC6-C3C chimeras. B, Western blot ( WB ) of lysates from HEK 293T cells transfected with BFP-TRPC3, BFP-TRPC6, BFP-TRPC6-C3C, BFP-TRPC6-C3N, or BFP-TRPC3-C6C. Blots were probed with antibodies that recognize the C terminus of human TRPC3 (anti-TRPC3 (C)), the N terminus of murine and human TRPC3 (anti-TRPC3 (N)), or the N terminus of TRPC6 (anti-TRPC6 (N)). Representative results of two experiments are shown.

    Article Snippet: Blots were incubated with anti-TRPC3 (1:400, Alomone Labs, Jerusalem, Israel), anti-TRPC3 (C) targeted to the human C-terminal sequence RRRRLQDIEMGMGNSKSRLN (1:1000, Bethyl Laboratories, Inc., Montgomery, TX) , anti-TRPC3 (N) targeted to the murine N-terminal sequence LNGDLESAEPLERHGHKASL (1:1000, Bethyl Laboratories, Inc.) , anti-TRPC6 (1:250, Alomone Labs; 1:500, Abcam, Inc., Cambridge, MA), anti-V5-HRP (1:10,000, Invitrogen), anti-FLAG (1:1000, Sigma), anti-PLCγ (1:1000, SC-81, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), anti-Epo-R (1:1000, SC697, Santa Cruz Biotechnology, Inc.), anti-actin (1:10,000, Sigma), anti-tubulin (1:10,000, Sigma) antibodies, or streptavidin-HRP (Pierce).

    Techniques: Western Blot, Transfection

    Interaction of TRPC3/TRPC6 chimeras with Epo-R. A, HEK 293T cells were transfected ( Tx'd ) with Epo-R and V5-TRPC3 or V5-TRPC3-C6C. Lysates were immunoprecipitated ( IP ) with anti-V5, anti-Epo-R antibodies, or normal rabbit serum ( NRS ). Western blots ( WB ) of lysates and immunoprecipitates were probed with anti-V5-HRP or anti-Epo-R and appropriate secondary antibody. Representative results of three similar experiments are shown. B, HEK 293T cells were transfected with Epo-R and FLAG-TRPC6, FLAG-TRPC6-C3C, or FLAG-TRPC6-C3N. Lysates were immunoprecipitated with anti-FLAG-agarose, anti-Epo-R, or normal rabbit serum. Western blots of lysates and immunoprecipitates were probed with anti-FLAG or anti-Epo-R antibodies. Representative results of three experiments are shown. C, densitometry was used to quantitate Epo-R, V5-TRPC3, FLAG-TRPC6, and chimeric channel bands from three experiments using transfected HEK 293T cells. The Epo-R to V5-TRPC3 or V5-TRPC3-C6C ratio or Epo-R to FLAG-TRPC6, FLAG-TRPC6-C3C, or FLAG-TRPC6-C3N ratio was calculated and normalized to V5-TRPC3 or FLAG-TRPC6 to allow comparison between experiments. The mean normalized ratio ± S.E. was determined for three separate experiments. The Epo-R/V5-TRPC3-C6C ratio was significantly less than the Epo-R/V5-TRPC3 ratio ( * , p ≤ 0.01), and the Epo-R/FLAG-TRPC6-C3C ratio was significantly greater than the Epo-R/FLAG-TRPC6 or FLAG-TRPC6-C3N ratio ( ** , *** , p ≤ 0.001).

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Activation by Erythropoietin Is Modulated by TRPC6

    doi: 10.1074/jbc.M804734200

    Figure Lengend Snippet: Interaction of TRPC3/TRPC6 chimeras with Epo-R. A, HEK 293T cells were transfected ( Tx'd ) with Epo-R and V5-TRPC3 or V5-TRPC3-C6C. Lysates were immunoprecipitated ( IP ) with anti-V5, anti-Epo-R antibodies, or normal rabbit serum ( NRS ). Western blots ( WB ) of lysates and immunoprecipitates were probed with anti-V5-HRP or anti-Epo-R and appropriate secondary antibody. Representative results of three similar experiments are shown. B, HEK 293T cells were transfected with Epo-R and FLAG-TRPC6, FLAG-TRPC6-C3C, or FLAG-TRPC6-C3N. Lysates were immunoprecipitated with anti-FLAG-agarose, anti-Epo-R, or normal rabbit serum. Western blots of lysates and immunoprecipitates were probed with anti-FLAG or anti-Epo-R antibodies. Representative results of three experiments are shown. C, densitometry was used to quantitate Epo-R, V5-TRPC3, FLAG-TRPC6, and chimeric channel bands from three experiments using transfected HEK 293T cells. The Epo-R to V5-TRPC3 or V5-TRPC3-C6C ratio or Epo-R to FLAG-TRPC6, FLAG-TRPC6-C3C, or FLAG-TRPC6-C3N ratio was calculated and normalized to V5-TRPC3 or FLAG-TRPC6 to allow comparison between experiments. The mean normalized ratio ± S.E. was determined for three separate experiments. The Epo-R/V5-TRPC3-C6C ratio was significantly less than the Epo-R/V5-TRPC3 ratio ( * , p ≤ 0.01), and the Epo-R/FLAG-TRPC6-C3C ratio was significantly greater than the Epo-R/FLAG-TRPC6 or FLAG-TRPC6-C3N ratio ( ** , *** , p ≤ 0.001).

    Article Snippet: Blots were incubated with anti-TRPC3 (1:400, Alomone Labs, Jerusalem, Israel), anti-TRPC3 (C) targeted to the human C-terminal sequence RRRRLQDIEMGMGNSKSRLN (1:1000, Bethyl Laboratories, Inc., Montgomery, TX) , anti-TRPC3 (N) targeted to the murine N-terminal sequence LNGDLESAEPLERHGHKASL (1:1000, Bethyl Laboratories, Inc.) , anti-TRPC6 (1:250, Alomone Labs; 1:500, Abcam, Inc., Cambridge, MA), anti-V5-HRP (1:10,000, Invitrogen), anti-FLAG (1:1000, Sigma), anti-PLCγ (1:1000, SC-81, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), anti-Epo-R (1:1000, SC697, Santa Cruz Biotechnology, Inc.), anti-actin (1:10,000, Sigma), anti-tubulin (1:10,000, Sigma) antibodies, or streptavidin-HRP (Pierce).

    Techniques: Transfection, Immunoprecipitation, Western Blot

    Schema of chimeras of subdivided TRPC3 and TRPC3 C termini. Amino acid ( AA ) composition of TRPC3 and TRPC6 chimeras and localization of TRP, CRIB, and coiled-coil domains are shown.

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Activation by Erythropoietin Is Modulated by TRPC6

    doi: 10.1074/jbc.M804734200

    Figure Lengend Snippet: Schema of chimeras of subdivided TRPC3 and TRPC3 C termini. Amino acid ( AA ) composition of TRPC3 and TRPC6 chimeras and localization of TRP, CRIB, and coiled-coil domains are shown.

    Article Snippet: Blots were incubated with anti-TRPC3 (1:400, Alomone Labs, Jerusalem, Israel), anti-TRPC3 (C) targeted to the human C-terminal sequence RRRRLQDIEMGMGNSKSRLN (1:1000, Bethyl Laboratories, Inc., Montgomery, TX) , anti-TRPC3 (N) targeted to the murine N-terminal sequence LNGDLESAEPLERHGHKASL (1:1000, Bethyl Laboratories, Inc.) , anti-TRPC6 (1:250, Alomone Labs; 1:500, Abcam, Inc., Cambridge, MA), anti-V5-HRP (1:10,000, Invitrogen), anti-FLAG (1:1000, Sigma), anti-PLCγ (1:1000, SC-81, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), anti-Epo-R (1:1000, SC697, Santa Cruz Biotechnology, Inc.), anti-actin (1:10,000, Sigma), anti-tubulin (1:10,000, Sigma) antibodies, or streptavidin-HRP (Pierce).

    Techniques:

    Immunolocalization of TRPC3 and Epo-R. HEK 293T cells were transfected ( Tx'd ) with Ext-V5-TRPC3, Epo-R, with or without TRPC6. Cells were permeabilized or not. Cells were stained with anti-V5, anti-Epo-R, or anti-TRPC6 antibodies, and then with Alexa Fluor 488 goat anti-mouse and Alexa Fluor 594 goat anti-rabbit antibodies. Nuclei were identified by 4′,6-diamidino-2-phenylindole staining. Representative results of images obtained in the midplane of the cell with confocal microscopy are shown.

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Activation by Erythropoietin Is Modulated by TRPC6

    doi: 10.1074/jbc.M804734200

    Figure Lengend Snippet: Immunolocalization of TRPC3 and Epo-R. HEK 293T cells were transfected ( Tx'd ) with Ext-V5-TRPC3, Epo-R, with or without TRPC6. Cells were permeabilized or not. Cells were stained with anti-V5, anti-Epo-R, or anti-TRPC6 antibodies, and then with Alexa Fluor 488 goat anti-mouse and Alexa Fluor 594 goat anti-rabbit antibodies. Nuclei were identified by 4′,6-diamidino-2-phenylindole staining. Representative results of images obtained in the midplane of the cell with confocal microscopy are shown.

    Article Snippet: Blots were incubated with anti-TRPC3 (1:400, Alomone Labs, Jerusalem, Israel), anti-TRPC3 (C) targeted to the human C-terminal sequence RRRRLQDIEMGMGNSKSRLN (1:1000, Bethyl Laboratories, Inc., Montgomery, TX) , anti-TRPC3 (N) targeted to the murine N-terminal sequence LNGDLESAEPLERHGHKASL (1:1000, Bethyl Laboratories, Inc.) , anti-TRPC6 (1:250, Alomone Labs; 1:500, Abcam, Inc., Cambridge, MA), anti-V5-HRP (1:10,000, Invitrogen), anti-FLAG (1:1000, Sigma), anti-PLCγ (1:1000, SC-81, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), anti-Epo-R (1:1000, SC697, Santa Cruz Biotechnology, Inc.), anti-actin (1:10,000, Sigma), anti-tubulin (1:10,000, Sigma) antibodies, or streptavidin-HRP (Pierce).

    Techniques: Transfection, Staining, Confocal Microscopy

    Modulation of membrane insertion of TRPC3 by Epo detected by cell surface biotinylation. HEK 293T cells transfected ( Tx'd ) with Epo-R and V5-TRPC3 without ( A and B ) or with FLAG-TRPC6 ( C and D ) were stimulated with 40 units/ml Epo. Biotinylation of cell surface proteins was performed, and V5-TRPC3 immunoprecipitated ( IP ) from lysates with anti-V5 antibody. Western blots ( WB ) were probed with streptavidin-HRP to detect biotinylated TRPC3 and anti-V5-HRP to detect total V5-TRPC3. Representative results of Western blots from four experiments are shown in A and C . Biotinylated and total TRPC3 bands were quantitated with densitometry, and the ratio was normalized to time 0. The mean ± S.E. of the biotinylated/total TRPC3 ratios at 0, 1, 5, 10, and 20 min from four experiments are shown ( B and D ). * indicates a significant difference in the ratio compared with time 0 ( p ≤ 0.02). E, Epo-stimulated cell surface expression of endogenous TRPC3 was examined using BFU-E-derived erythroblasts at day 10 of methylcellulose culture (two experiments) or erythroblasts from phase II day 8 of liquid culture (one experiment). Cells were stimulated with 40 units/ml Epo for 0 or 5 min and biotinylated, and TRPC3 was immunoprecipitated from lysates with anti-TRPC3 antibody. Western blots were probed with streptavidin-HRP to detect biotinylated TRPC3 and anti-TRPC3 to detect total TRPC3. A representative result of three Western blots is shown. F, biotinylated and total TRPC3 bands were quantitated with densitometry, and the ratio was normalized to time 0. The mean ± S.E. of the biotinylated/total TRPC3 ratios at 0 and 5 min from the three experiments are shown. No significant difference in the ratio at 5 min compared with time 0 was detected.

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Activation by Erythropoietin Is Modulated by TRPC6

    doi: 10.1074/jbc.M804734200

    Figure Lengend Snippet: Modulation of membrane insertion of TRPC3 by Epo detected by cell surface biotinylation. HEK 293T cells transfected ( Tx'd ) with Epo-R and V5-TRPC3 without ( A and B ) or with FLAG-TRPC6 ( C and D ) were stimulated with 40 units/ml Epo. Biotinylation of cell surface proteins was performed, and V5-TRPC3 immunoprecipitated ( IP ) from lysates with anti-V5 antibody. Western blots ( WB ) were probed with streptavidin-HRP to detect biotinylated TRPC3 and anti-V5-HRP to detect total V5-TRPC3. Representative results of Western blots from four experiments are shown in A and C . Biotinylated and total TRPC3 bands were quantitated with densitometry, and the ratio was normalized to time 0. The mean ± S.E. of the biotinylated/total TRPC3 ratios at 0, 1, 5, 10, and 20 min from four experiments are shown ( B and D ). * indicates a significant difference in the ratio compared with time 0 ( p ≤ 0.02). E, Epo-stimulated cell surface expression of endogenous TRPC3 was examined using BFU-E-derived erythroblasts at day 10 of methylcellulose culture (two experiments) or erythroblasts from phase II day 8 of liquid culture (one experiment). Cells were stimulated with 40 units/ml Epo for 0 or 5 min and biotinylated, and TRPC3 was immunoprecipitated from lysates with anti-TRPC3 antibody. Western blots were probed with streptavidin-HRP to detect biotinylated TRPC3 and anti-TRPC3 to detect total TRPC3. A representative result of three Western blots is shown. F, biotinylated and total TRPC3 bands were quantitated with densitometry, and the ratio was normalized to time 0. The mean ± S.E. of the biotinylated/total TRPC3 ratios at 0 and 5 min from the three experiments are shown. No significant difference in the ratio at 5 min compared with time 0 was detected.

    Article Snippet: Blots were incubated with anti-TRPC3 (1:400, Alomone Labs, Jerusalem, Israel), anti-TRPC3 (C) targeted to the human C-terminal sequence RRRRLQDIEMGMGNSKSRLN (1:1000, Bethyl Laboratories, Inc., Montgomery, TX) , anti-TRPC3 (N) targeted to the murine N-terminal sequence LNGDLESAEPLERHGHKASL (1:1000, Bethyl Laboratories, Inc.) , anti-TRPC6 (1:250, Alomone Labs; 1:500, Abcam, Inc., Cambridge, MA), anti-V5-HRP (1:10,000, Invitrogen), anti-FLAG (1:1000, Sigma), anti-PLCγ (1:1000, SC-81, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), anti-Epo-R (1:1000, SC697, Santa Cruz Biotechnology, Inc.), anti-actin (1:10,000, Sigma), anti-tubulin (1:10,000, Sigma) antibodies, or streptavidin-HRP (Pierce).

    Techniques: Transfection, Immunoprecipitation, Western Blot, Expressing, Derivative Assay

    Plasma membrane insertion of TRPC3/TRPC3 chimeras detected with cell surface biotinylation. Cell surface biotinylation was performed with HEK 293T cells expressing V5-TRPC3, V5-TRPC3-C6C, V5-TRPC3-C6C1, V5-TRPC3-C6C2, FLAG-TRPC6, FLAG-TRPC6-C3C, or FLAG-TRPC6-C3N and Epo-R. Lysates were prepared, and immunoprecipitation ( IP ) performed with anti-V5 antibody or anti-FLAG-agarose. Western blotting ( WB ) was performed on immunoprecipitation pellets with streptavidin-HRP to detect biotinylation and either anti-V5-HRP to detect total TRPC3 chimeras or anti-TRPC6 or anti-TRPC3-N antibodies to detect total TRPC6 chimeras. Representative results of two experiments are shown. Tx'd , transfected.

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Activation by Erythropoietin Is Modulated by TRPC6

    doi: 10.1074/jbc.M804734200

    Figure Lengend Snippet: Plasma membrane insertion of TRPC3/TRPC3 chimeras detected with cell surface biotinylation. Cell surface biotinylation was performed with HEK 293T cells expressing V5-TRPC3, V5-TRPC3-C6C, V5-TRPC3-C6C1, V5-TRPC3-C6C2, FLAG-TRPC6, FLAG-TRPC6-C3C, or FLAG-TRPC6-C3N and Epo-R. Lysates were prepared, and immunoprecipitation ( IP ) performed with anti-V5 antibody or anti-FLAG-agarose. Western blotting ( WB ) was performed on immunoprecipitation pellets with streptavidin-HRP to detect biotinylation and either anti-V5-HRP to detect total TRPC3 chimeras or anti-TRPC6 or anti-TRPC3-N antibodies to detect total TRPC6 chimeras. Representative results of two experiments are shown. Tx'd , transfected.

    Article Snippet: Blots were incubated with anti-TRPC3 (1:400, Alomone Labs, Jerusalem, Israel), anti-TRPC3 (C) targeted to the human C-terminal sequence RRRRLQDIEMGMGNSKSRLN (1:1000, Bethyl Laboratories, Inc., Montgomery, TX) , anti-TRPC3 (N) targeted to the murine N-terminal sequence LNGDLESAEPLERHGHKASL (1:1000, Bethyl Laboratories, Inc.) , anti-TRPC6 (1:250, Alomone Labs; 1:500, Abcam, Inc., Cambridge, MA), anti-V5-HRP (1:10,000, Invitrogen), anti-FLAG (1:1000, Sigma), anti-PLCγ (1:1000, SC-81, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), anti-Epo-R (1:1000, SC697, Santa Cruz Biotechnology, Inc.), anti-actin (1:10,000, Sigma), anti-tubulin (1:10,000, Sigma) antibodies, or streptavidin-HRP (Pierce).

    Techniques: Expressing, Immunoprecipitation, Western Blot, Transfection

    The correlation between the plasma level of the total alternative renin angiotensin system activity (Alt-S) and the weight of the visceral fat ( a ) and body weight ( b ). The correlation between the plasma level of angiotensin 1–7 (Ang 1–7) and the weight of the visceral fat ( c ) and body weight ( d ) in SHRs and SHRs treated with MLN-4760 (SHR + MLN).

    Journal: Biomedicines

    Article Title: Vascular Effects of Low-Dose ACE2 Inhibitor MLN-4760—Benefit or Detriment in Essential Hypertension?

    doi: 10.3390/biomedicines10010038

    Figure Lengend Snippet: The correlation between the plasma level of the total alternative renin angiotensin system activity (Alt-S) and the weight of the visceral fat ( a ) and body weight ( b ). The correlation between the plasma level of angiotensin 1–7 (Ang 1–7) and the weight of the visceral fat ( c ) and body weight ( d ) in SHRs and SHRs treated with MLN-4760 (SHR + MLN).

    Article Snippet: Membranes were blocked with 5% nonfat milk for 1 h at room temperature in Tris-buffer solution (pH 7.6) containing 0.1% Tween-20 and probed against the following primary antibodies: rabbit polyclonal anti-endothelial NOS, anti-neuronal NOS and anti-β-actin (Abcam, Cambridge, UK); rabbit polyclonal anti-inducible NOS (Bio-Rad, Inc., Hercules, CA, USA) rabbit polyclonal anti-angiotensin(1–7) Mas receptor (Alomone Labs, Jerusalem, Israel); rabbit polyclonal CBS and mouse monoclonal anti-CSE antibodies (Proteintech, Manchester, UK); and rabbit monoclonal anti-ACE2 (Invitrogen, Waltham, MA, USA) overnight at 4 °C.

    Techniques: Activity Assay

    Immunofluorescence studies in HEK293T cells overexpressing c-myc tagged MasR using MasR antibodies. Images of fluorescence signal corresponding to secondary cyanine (Cy3)-conjugated antibody (A), nuclear Hoechst 33258 staining (B), merged images (C) and bright field (D) are shown. In cells transfected with the pcDNA 3.1/c-myc-MasR construct, antibodies NLS-1531 and sc-54682 generated a similar staining pattern to that generated with the anti c-myc antibody. No staining was observed with any of these antibodies in cells transfected with the empty vector pcDNA 3.1. Antibody sc-135063 was capable of staining the plasma membrane of cells overexpressing the MasR but also generated widespread signals in cells transfected with the empty vector. For antibody AAR-013 there was no staining associated with the cell membrane. Intense immunocytochemical staining of identical distribution and intensity was revealed in cells transfected with the empty vector and those transfected with the c-myc-MasR construct. Images are representative of 3 independent experiments.

    Journal: PLoS ONE

    Article Title: Validation of commercial Mas receptor antibodies for utilization in Western Blotting, immunofluorescence and immunohistochemistry studies

    doi: 10.1371/journal.pone.0183278

    Figure Lengend Snippet: Immunofluorescence studies in HEK293T cells overexpressing c-myc tagged MasR using MasR antibodies. Images of fluorescence signal corresponding to secondary cyanine (Cy3)-conjugated antibody (A), nuclear Hoechst 33258 staining (B), merged images (C) and bright field (D) are shown. In cells transfected with the pcDNA 3.1/c-myc-MasR construct, antibodies NLS-1531 and sc-54682 generated a similar staining pattern to that generated with the anti c-myc antibody. No staining was observed with any of these antibodies in cells transfected with the empty vector pcDNA 3.1. Antibody sc-135063 was capable of staining the plasma membrane of cells overexpressing the MasR but also generated widespread signals in cells transfected with the empty vector. For antibody AAR-013 there was no staining associated with the cell membrane. Intense immunocytochemical staining of identical distribution and intensity was revealed in cells transfected with the empty vector and those transfected with the c-myc-MasR construct. Images are representative of 3 independent experiments.

    Article Snippet: Then cells were incubated for 1 h at room temperature with the anti-c-myc antibody (1:200) or with four different anti-MasR antibodies that were recommended for immunocytochemistry [NLS-1531, sc-135063 and sc-54682, (diluted 1:200) and AAR-013 (diluted 1:100)].

    Techniques: Immunofluorescence, Fluorescence, Staining, Transfection, Construct, Generated, Plasmid Preparation

    Western blotting of HEK293T cells overexpressing c-myc tagged MasR using anti c-myc tag and three MasR antibodies. Cells were transfected with varying amounts of a plasmid encoding the c-myc-tagged MasR (0.2 and 0.5 μg of pcDNA 3.1/c-myc-MasR).The anti-c-myc antibody detected a band of 37 kDa whose intensity increased as higher amount of DNA was transfected (arrow). Additionally, a ladder of higher molecular weight (MW) immunoreactive bands could be observed (asterisk). sc-54682 antibody reproduced the band pattern obtained with the c-myc antibody (arrow and asterisk). sc-135063 antibody revealed multiple bands close to 37 kDa with one of them of increasing intensity as higher amount of DNA was transfected (arrow) but did not produce a ladder of higher MW immunoreactive bands. High unspecific staining was attained with sc-135063 antibody. AAR-013 antibody revealed a pattern of immunoreactivity not comparable to those obtained with the other antibodies. Detection of β-tubulin was used as a protein loading control. The membranes are representative of 3 independent experiments.

    Journal: PLoS ONE

    Article Title: Validation of commercial Mas receptor antibodies for utilization in Western Blotting, immunofluorescence and immunohistochemistry studies

    doi: 10.1371/journal.pone.0183278

    Figure Lengend Snippet: Western blotting of HEK293T cells overexpressing c-myc tagged MasR using anti c-myc tag and three MasR antibodies. Cells were transfected with varying amounts of a plasmid encoding the c-myc-tagged MasR (0.2 and 0.5 μg of pcDNA 3.1/c-myc-MasR).The anti-c-myc antibody detected a band of 37 kDa whose intensity increased as higher amount of DNA was transfected (arrow). Additionally, a ladder of higher molecular weight (MW) immunoreactive bands could be observed (asterisk). sc-54682 antibody reproduced the band pattern obtained with the c-myc antibody (arrow and asterisk). sc-135063 antibody revealed multiple bands close to 37 kDa with one of them of increasing intensity as higher amount of DNA was transfected (arrow) but did not produce a ladder of higher MW immunoreactive bands. High unspecific staining was attained with sc-135063 antibody. AAR-013 antibody revealed a pattern of immunoreactivity not comparable to those obtained with the other antibodies. Detection of β-tubulin was used as a protein loading control. The membranes are representative of 3 independent experiments.

    Article Snippet: Then cells were incubated for 1 h at room temperature with the anti-c-myc antibody (1:200) or with four different anti-MasR antibodies that were recommended for immunocytochemistry [NLS-1531, sc-135063 and sc-54682, (diluted 1:200) and AAR-013 (diluted 1:100)].

    Techniques: Western Blot, Transfection, Plasmid Preparation, Molecular Weight, Staining

    Immunohistochemistry in heart and kidney from wild-type (WT) and MasR-KO mice. Negative controls performed by omitting the primary antibody demonstrated minimal background immunostaining in heart (A) and kidney (B) sections. (A) In heart sections, NLS-1531 antibody stained predominantly the cytoplasm of cardiomyocytes with similar intensity in WT and MasR-KO hearts. The AAR-013 antibody stained the cardiomyocytes nucleus and weaker staining was observed in their cytoplasm. The same pattern was found in heart sections of MasR-KO mice. (B) In kidney sections, for NLS-1531 antibody, staining was mostly restricted to cytoplasm of numerous tubules cells with similar intensity in WT and MasR-KO kidneys. Antibody AAR-013 stained most intensely tubules cells nucleus and weaker staining was observed in their cytoplasm. Glomeruli were also stained positively. The same pattern was found in kidney sections of MasR-KO. Preincubation of the AAR-013 antibody with the blocking peptide provided by the vendors eliminated the immunohistochemical nuclear staining both in WT and MasR-KO mice kidney sections. Images are representative of n = 3. Bar, 50 μm.

    Journal: PLoS ONE

    Article Title: Validation of commercial Mas receptor antibodies for utilization in Western Blotting, immunofluorescence and immunohistochemistry studies

    doi: 10.1371/journal.pone.0183278

    Figure Lengend Snippet: Immunohistochemistry in heart and kidney from wild-type (WT) and MasR-KO mice. Negative controls performed by omitting the primary antibody demonstrated minimal background immunostaining in heart (A) and kidney (B) sections. (A) In heart sections, NLS-1531 antibody stained predominantly the cytoplasm of cardiomyocytes with similar intensity in WT and MasR-KO hearts. The AAR-013 antibody stained the cardiomyocytes nucleus and weaker staining was observed in their cytoplasm. The same pattern was found in heart sections of MasR-KO mice. (B) In kidney sections, for NLS-1531 antibody, staining was mostly restricted to cytoplasm of numerous tubules cells with similar intensity in WT and MasR-KO kidneys. Antibody AAR-013 stained most intensely tubules cells nucleus and weaker staining was observed in their cytoplasm. Glomeruli were also stained positively. The same pattern was found in kidney sections of MasR-KO. Preincubation of the AAR-013 antibody with the blocking peptide provided by the vendors eliminated the immunohistochemical nuclear staining both in WT and MasR-KO mice kidney sections. Images are representative of n = 3. Bar, 50 μm.

    Article Snippet: Then cells were incubated for 1 h at room temperature with the anti-c-myc antibody (1:200) or with four different anti-MasR antibodies that were recommended for immunocytochemistry [NLS-1531, sc-135063 and sc-54682, (diluted 1:200) and AAR-013 (diluted 1:100)].

    Techniques: Immunohistochemistry, Mouse Assay, Immunostaining, Staining, Blocking Assay