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Effect of <t>B1R</t> activation on ADAM17 activity in primary hypothalamic neurons. ( A ) Incubation of wild-type neurons with B1R-specific agonist Lys-Des-Arg 9 -Bradykinin (LDABK) significantly increased ADAM17 activity. Pre-treatment with R715, a B1R-specific antagonist, prevented this LDABK-mediated increase in ADAM17 activity. R715 treatment alone did not have any effect on ADAM17 activity. ( B ) Treatment with LDABK did not show any effect on ADAM17 activity in neurons with B1R deletion. In addition, the HOE 140, a kinin B2 receptor (B2R)-specific antagonist also did not show any effect on ADAM17 activity. The cultured primary neurons were pre-treated with a specific B1R antagonist (R715, 10 μM) or a specific B2R antagonist HOE 140 (HOE, 10 μM) for 1 h, followed by treatment with LDABK (300 nM) for 18 h ( n = 6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

B1r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/b1r/product/Alomone Labs
Average 94 stars, based on 2 article reviews
Price from $9.99 to $1999.99

b1r - by Bioz Stars, 2022-08

94/100 stars

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1) Product Images from "Activation of Kinin B1R Upregulates ADAM17 and Results in ACE2 Shedding in Neurons"

Article Title: Activation of Kinin B1R Upregulates ADAM17 and Results in ACE2 Shedding in Neurons

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms22010145

Effect of B1R activation on ADAM17 activity in primary hypothalamic neurons. ( A ) Incubation of wild-type neurons with B1R-specific agonist Lys-Des-Arg 9 -Bradykinin (LDABK) significantly increased ADAM17 activity. Pre-treatment with R715, a B1R-specific antagonist, prevented this LDABK-mediated increase in ADAM17 activity. R715 treatment alone did not have any effect on ADAM17 activity. ( B ) Treatment with LDABK did not show any effect on ADAM17 activity in neurons with B1R deletion. In addition, the HOE 140, a kinin B2 receptor (B2R)-specific antagonist also did not show any effect on ADAM17 activity. The cultured primary neurons were pre-treated with a specific B1R antagonist (R715, 10 μM) or a specific B2R antagonist HOE 140 (HOE, 10 μM) for 1 h, followed by treatment with LDABK (300 nM) for 18 h ( n = 6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Figure Legend Snippet: Effect of B1R activation on ADAM17 activity in primary hypothalamic neurons. ( A ) Incubation of wild-type neurons with B1R-specific agonist Lys-Des-Arg 9 -Bradykinin (LDABK) significantly increased ADAM17 activity. Pre-treatment with R715, a B1R-specific antagonist, prevented this LDABK-mediated increase in ADAM17 activity. R715 treatment alone did not have any effect on ADAM17 activity. ( B ) Treatment with LDABK did not show any effect on ADAM17 activity in neurons with B1R deletion. In addition, the HOE 140, a kinin B2 receptor (B2R)-specific antagonist also did not show any effect on ADAM17 activity. The cultured primary neurons were pre-treated with a specific B1R antagonist (R715, 10 μM) or a specific B2R antagonist HOE 140 (HOE, 10 μM) for 1 h, followed by treatment with LDABK (300 nM) for 18 h ( n = 6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques Used: Activation Assay, Activity Assay, Incubation, Cell Culture

Effect of B1R activation on ACE2 activity in primary hypothalamic neurons. ( A ) Incubation of wild-type neurons with B1R-specific agonist LDABK significantly decreased ACE2 activity. R715, a B1R-specific antagonist pretreatment prevented this LDABK-mediated decrease in ACE2 activity. R715 treatment alone did not have any effect on ACE2 activity. ( B ) Treatment with LDABK did not show any effect on ACE2 activity in neurons with B1R deletion. In addition, the HOE 140, a B2R-specific antagonist also did not show any effect on ACE2 activity. The cultured primary neurons were pre-treated with a specific B1R antagonist (R715, 10 μM) or a specific B2R antagonist HOE 140 (HOE, 10 μM) for 1 h, followed by treatment with LDABK (300 nM) for 18 h ( n = 6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Figure Legend Snippet: Effect of B1R activation on ACE2 activity in primary hypothalamic neurons. ( A ) Incubation of wild-type neurons with B1R-specific agonist LDABK significantly decreased ACE2 activity. R715, a B1R-specific antagonist pretreatment prevented this LDABK-mediated decrease in ACE2 activity. R715 treatment alone did not have any effect on ACE2 activity. ( B ) Treatment with LDABK did not show any effect on ACE2 activity in neurons with B1R deletion. In addition, the HOE 140, a B2R-specific antagonist also did not show any effect on ACE2 activity. The cultured primary neurons were pre-treated with a specific B1R antagonist (R715, 10 μM) or a specific B2R antagonist HOE 140 (HOE, 10 μM) for 1 h, followed by treatment with LDABK (300 nM) for 18 h ( n = 6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques Used: Activation Assay, Activity Assay, Incubation, Cell Culture

Effect of glutamate on B1R activation in primary hypothalamic neurons. Representative triple immunostaining revealed that compared to vehicle ( A ) treatment, stimulation with glutamate (100 μM) ( B ) for 18 h induced the expression of kinin B1R and ADAM17 protein in primary hypothalamic neurons. This glutamate-induced increase in B1R and ADAM17 protein expression was prevented by pre-treatment with R715 ( C ). Treatment with glutamate (100 μM) for 4 h induced an increase in B1R mRNA in cultured neurons, measured by real time PCR. This increase in B1R mRNA was prevented by pre-treatment with R715 ( D ). Representative image of Western blot ( E ) and quantitative analysis ( F ) showing glutamate stimulation induced B1R protein expression which was prevented by pre-treatment with R715 in neurons ( n = 4–6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Figure Legend Snippet: Effect of glutamate on B1R activation in primary hypothalamic neurons. Representative triple immunostaining revealed that compared to vehicle ( A ) treatment, stimulation with glutamate (100 μM) ( B ) for 18 h induced the expression of kinin B1R and ADAM17 protein in primary hypothalamic neurons. This glutamate-induced increase in B1R and ADAM17 protein expression was prevented by pre-treatment with R715 ( C ). Treatment with glutamate (100 μM) for 4 h induced an increase in B1R mRNA in cultured neurons, measured by real time PCR. This increase in B1R mRNA was prevented by pre-treatment with R715 ( D ). Representative image of Western blot ( E ) and quantitative analysis ( F ) showing glutamate stimulation induced B1R protein expression which was prevented by pre-treatment with R715 in neurons ( n = 4–6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques Used: Activation Assay, Triple Immunostaining, Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot

Kinin B1 receptor (B1R), A Disintegrin And Metalloprotease 17 (ADAM17), and angiotensin converting enzyme 2 (ACE2) expression in primary hypothalamic neurons. Representative photomicrographs showing immunofluorescence staining for neuron-specific marker microtubule associated protein 2 (MAP-2) (Green) in primary neurons from wild-type ( A ) and B1R knockout mice ( B ) cultured for 10 days. Primary neurons from wild-type mice pups showed immunopositivity for the presence of B1R ( C ), while B1R immunoreactivity was absent in neurons with B1R deletion ( D ). Representative triple immunostaining revealed that kinin B1R ( E ) and ADAM17 ( F ) are expressed in primary hypothalamic neurons expressing nuclear DAPI staining in blue ( G ), and merged image ( H ) shows the colocalization of B1R and ADAM17. Representative triple immunostaining showing ACE2 and ADAM17 expression and co-localization in wild-type neurons ( I – L ) and in B1R knockout neurons ( M – P ).

Figure Legend Snippet: Kinin B1 receptor (B1R), A Disintegrin And Metalloprotease 17 (ADAM17), and angiotensin converting enzyme 2 (ACE2) expression in primary hypothalamic neurons. Representative photomicrographs showing immunofluorescence staining for neuron-specific marker microtubule associated protein 2 (MAP-2) (Green) in primary neurons from wild-type ( A ) and B1R knockout mice ( B ) cultured for 10 days. Primary neurons from wild-type mice pups showed immunopositivity for the presence of B1R ( C ), while B1R immunoreactivity was absent in neurons with B1R deletion ( D ). Representative triple immunostaining revealed that kinin B1R ( E ) and ADAM17 ( F ) are expressed in primary hypothalamic neurons expressing nuclear DAPI staining in blue ( G ), and merged image ( H ) shows the colocalization of B1R and ADAM17. Representative triple immunostaining showing ACE2 and ADAM17 expression and co-localization in wild-type neurons ( I – L ) and in B1R knockout neurons ( M – P ).

Techniques Used: Expressing, Immunofluorescence, Staining, Marker, Knock-Out, Mouse Assay, Cell Culture, Triple Immunostaining

Activation of kinin B1R upregulates ADAM17-mediated ACE2 shedding in primary hypothalamic neurons. B1R activation by specific agonist results in upregulation of ADAM17 expression and activity which results in increased membrane-bound ACE2 shedding and reduced ACE2 activity in primary hypothalamic neurons. In addition, glutamate can induce B1R expression and B1R is involved in glutamate-mediated effects on ADAM17 activity and ACE2 shedding.

Figure Legend Snippet: Activation of kinin B1R upregulates ADAM17-mediated ACE2 shedding in primary hypothalamic neurons. B1R activation by specific agonist results in upregulation of ADAM17 expression and activity which results in increased membrane-bound ACE2 shedding and reduced ACE2 activity in primary hypothalamic neurons. In addition, glutamate can induce B1R expression and B1R is involved in glutamate-mediated effects on ADAM17 activity and ACE2 shedding.

Techniques Used: Activation Assay, Expressing, Activity Assay

Glutamate-induced ADAM17-mediated ACE2 shedding involves kinin B1R activation. Glutamate increased ADAM17 activity ( A ) and decreased ACE2 activity ( B ) in neurons, which was prevented by pre-treatment with R715. Treatment of B1R knockout neurons with glutamate did not show any effect on ADAM17 or ACE2 activity. ADAM17 and ACE2 assays were performed in primary hypothalamic neurons cultured from wild-type and B1R knockout neonates, treated with glutamate (Glut, 100 μM) or glutamate with 1 h of pre-treatment with R715 (a specific B1R inhibitor, 10 μM) for 18 h ( n = 6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Figure Legend Snippet: Glutamate-induced ADAM17-mediated ACE2 shedding involves kinin B1R activation. Glutamate increased ADAM17 activity ( A ) and decreased ACE2 activity ( B ) in neurons, which was prevented by pre-treatment with R715. Treatment of B1R knockout neurons with glutamate did not show any effect on ADAM17 or ACE2 activity. ADAM17 and ACE2 assays were performed in primary hypothalamic neurons cultured from wild-type and B1R knockout neonates, treated with glutamate (Glut, 100 μM) or glutamate with 1 h of pre-treatment with R715 (a specific B1R inhibitor, 10 μM) for 18 h ( n = 6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques Used: Activation Assay, Activity Assay, Knock-Out, Cell Culture

2) Product Images from "Kinin B1 Receptor Blockade Prevents Angiotensin II-induced Neuroinflammation and Oxidative Stress in Primary Hypothalamic Neurons"

Article Title: Kinin B1 Receptor Blockade Prevents Angiotensin II-induced Neuroinflammation and Oxidative Stress in Primary Hypothalamic Neurons

Journal: Cellular and molecular neurobiology

doi: 10.1007/s10571-019-00778-1

B1R antagonist treatment reduced angiotensin II induced NF-κB binding activity in primary hypothalamic neurons. The cultured primary neurons are pre-treated with B1R specific antagonist (R715, 10 μM) for 1 hour, followed by treatment with vehicle or angiotensin II (Ang II, 300 nM) for 24 hours. NF-kB DNA binding activity was analyzed by NF-kB p65 transcription factor assay kit. The result of NF-kB DNA binding showed that Ang II treatment significant increased NF-kB p65 DNA binding activity in primary neurons, and treatment with R715 significantly attenuated this effect. (n=6 independent culture wells/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p

Figure Legend Snippet: B1R antagonist treatment reduced angiotensin II induced NF-κB binding activity in primary hypothalamic neurons. The cultured primary neurons are pre-treated with B1R specific antagonist (R715, 10 μM) for 1 hour, followed by treatment with vehicle or angiotensin II (Ang II, 300 nM) for 24 hours. NF-kB DNA binding activity was analyzed by NF-kB p65 transcription factor assay kit. The result of NF-kB DNA binding showed that Ang II treatment significant increased NF-kB p65 DNA binding activity in primary neurons, and treatment with R715 significantly attenuated this effect. (n=6 independent culture wells/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques Used: Binding Assay, Activity Assay, Cell Culture, Transcription Factor Assay

Figure Legend Snippet: B1R antagonist treatment prevented angiotensin II-induced decrease in mitochondrial respiration in primary hypothalamic neurons. Primary hypothalamic mouse neurons were cultured on Seahorse XF-24 plates and treated with vehicle or Ang II with or without R715 for 24 hours. Seahorse mito stress assay was performed to measure oxygen consumption rate (OCR). (A). Bioenergetic profile following sequential injection of oligomycin ( O ), FCCP ( F ) and antimycin A/rotenone ( A/R ) indicating the key parameters of mitochondrial respiration in neurons. OCR measurements showed that Ang II stimulation resulted in significant decrease in basal respiration (B), ATP production (C), maximal respiration (D), and spare respiratory capacity (E) indicating mitochondrial dysfunction, and treatment with R715 prevented this Ang II-induced mitochondrial dysfunction. (n=8 independent culture wells/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques Used: Cell Culture, Injection

Angiotensin II-induced neuroinflammation and oxidative stress are mediated by kinin B1R. Mouse neonatal primary hypothermic neurons express both B1R and AT1R. Ang II stimulation of neurons increased expression of B1R, which in turn increased resulted in upregulation of Nox2 and Nox4 expression, and NF-κB activation, and ultimately leading to expression of pro-inflammatory cytokine production. Treatment with R715, the specific B1R antagonist blunted angiotensin II-induced neuroinflammation and oxidative stress by reducing Nox gene expression and attenuating NF-κB activation.

Figure Legend Snippet: Angiotensin II-induced neuroinflammation and oxidative stress are mediated by kinin B1R. Mouse neonatal primary hypothermic neurons express both B1R and AT1R. Ang II stimulation of neurons increased expression of B1R, which in turn increased resulted in upregulation of Nox2 and Nox4 expression, and NF-κB activation, and ultimately leading to expression of pro-inflammatory cytokine production. Treatment with R715, the specific B1R antagonist blunted angiotensin II-induced neuroinflammation and oxidative stress by reducing Nox gene expression and attenuating NF-κB activation.

Techniques Used: Expressing, Activation Assay

Angiotensin II-induced inflammatory gene expression is reduced by treatment with B1R antagonist in primary hypothalamic neurons. Angiotensin II treatment induced neuroinflammation as indicated by increase in pro-inflammatory genes (A) cyclooxygenase (Cox2), (B) Interleukin (IL)-1β, (C) IL-6, and (D) tumor necrosis factor (TNF). This increase in neuroinflammation was prevented by pre-treatment with R715. The cultured primary neurons are pre-treated with a specific B1R antagonist (R715, 10 μM) for 1 hour, followed by treatment with angiotensin II (Ang II, 300 nM) for 6 hours. The gene expression was measured using real time RT-PCR in triplicates. (n=4 independent cultures/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p

Figure Legend Snippet: Angiotensin II-induced inflammatory gene expression is reduced by treatment with B1R antagonist in primary hypothalamic neurons. Angiotensin II treatment induced neuroinflammation as indicated by increase in pro-inflammatory genes (A) cyclooxygenase (Cox2), (B) Interleukin (IL)-1β, (C) IL-6, and (D) tumor necrosis factor (TNF). This increase in neuroinflammation was prevented by pre-treatment with R715. The cultured primary neurons are pre-treated with a specific B1R antagonist (R715, 10 μM) for 1 hour, followed by treatment with angiotensin II (Ang II, 300 nM) for 6 hours. The gene expression was measured using real time RT-PCR in triplicates. (n=4 independent cultures/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques Used: Expressing, Cell Culture, Quantitative RT-PCR

Kinin B1 receptor gene expression is induced by angiotensin II in primary hypothalamic neurons. Brightfield photomicrograph showing primary mouse hypothalamic neuron cultures grown for 5 days (A). Representative photomicrographs showing immunofluorescence staining for neuron specific marker microtubule associated protein 2, MAP-2 (Red) and glial cell specific marker glial fibrillary acidic protein, GFAP (Green) in primary neurons cultured for 10 days without Ara-C (B) and with Ara-C (C) treatment. Treatment with Ara-C for 14 days resulted in primarily predominant neuronal population as stained for neuronal marker MAP-2 (D). Triple immunostaining revealed that Kinin B1R (E) and AT1R (F) are expressed in primary hypothalamic neurons. Treatment with angiotensin II (300 nM) induced increase in B1R mRNA levels in cultured neurons, measured by real time PCR (G). (n=4 independent cultures/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p

Figure Legend Snippet: Kinin B1 receptor gene expression is induced by angiotensin II in primary hypothalamic neurons. Brightfield photomicrograph showing primary mouse hypothalamic neuron cultures grown for 5 days (A). Representative photomicrographs showing immunofluorescence staining for neuron specific marker microtubule associated protein 2, MAP-2 (Red) and glial cell specific marker glial fibrillary acidic protein, GFAP (Green) in primary neurons cultured for 10 days without Ara-C (B) and with Ara-C (C) treatment. Treatment with Ara-C for 14 days resulted in primarily predominant neuronal population as stained for neuronal marker MAP-2 (D). Triple immunostaining revealed that Kinin B1R (E) and AT1R (F) are expressed in primary hypothalamic neurons. Treatment with angiotensin II (300 nM) induced increase in B1R mRNA levels in cultured neurons, measured by real time PCR (G). (n=4 independent cultures/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques Used: Expressing, Immunofluorescence, Staining, Marker, Cell Culture, Acetylene Reduction Assay, Triple Immunostaining, Real-time Polymerase Chain Reaction

B1R antagonist treatment reduced angiotensin II induced reactive oxygen species (ROS) production in primary hypothalamic neurons. (A). Representative photomicrographs showing dihydroethidium (DHE) stained primary hypothalamic neurons. (B). DHE staining quantified as corrected total cell fluorescence shows increased superoxide generation by treatment with angiotensin II (Ang II), which was attenuated by R715 treatment. (n=8/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p

Figure Legend Snippet: B1R antagonist treatment reduced angiotensin II induced reactive oxygen species (ROS) production in primary hypothalamic neurons. (A). Representative photomicrographs showing dihydroethidium (DHE) stained primary hypothalamic neurons. (B). DHE staining quantified as corrected total cell fluorescence shows increased superoxide generation by treatment with angiotensin II (Ang II), which was attenuated by R715 treatment. (n=8/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques Used: Staining, Fluorescence

Angiotensin II-induced Nox2 and Nox4 gene expression was reduced by treatment with B1R antagonist in primary hypothalamic neurons. Angiotensin II treatment induced oxidative stress by increased gene expression of (A) Nox2 and (B) Nox4. This increase was attenuated or prevented by pre-treatment with R715. The cultured primary neurons are pre-treated with a specific B1R antagonist (R715, 10 μM) for 1 hour, followed by treatment with angiotensin II (Ang II, 300 nM) for 6 hours. The gene expression was measured using real time RT-PCR in triplicates. (n=4 independent cultures/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p

Figure Legend Snippet: Angiotensin II-induced Nox2 and Nox4 gene expression was reduced by treatment with B1R antagonist in primary hypothalamic neurons. Angiotensin II treatment induced oxidative stress by increased gene expression of (A) Nox2 and (B) Nox4. This increase was attenuated or prevented by pre-treatment with R715. The cultured primary neurons are pre-treated with a specific B1R antagonist (R715, 10 μM) for 1 hour, followed by treatment with angiotensin II (Ang II, 300 nM) for 6 hours. The gene expression was measured using real time RT-PCR in triplicates. (n=4 independent cultures/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques Used: Expressing, Cell Culture, Quantitative RT-PCR

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anti b1 bradykinin receptor bdkrb1 antibody (Alomone Labs)

Alomone Labs anti b1 bradykinin receptor bdkrb1 antibody

<t>B1R</t> gene deletion prevents inflammation in the kidney during DOCA-salt-induced hypertension. DOCA-salt treatment for 3 weeks significantly increased mRNA expression of pro-inflammatory cytokines TNF, IL-6, IL-1β and chemokine MCP-1 in the kidney cortex of wild-type (WT) mice compared with sham mice. B1R knockout (B1RKO) mice did not show this DOCA-salt-induced increase in inflammatory markers. Gene expression measured by real time RT-PCR. Data was normalized to β-actin and presented as mean ± SEM ( n = 6, Two-way ANOVA, * P

Anti B1 Bradykinin Receptor Bdkrb1 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 b1 bradykinin receptor bdkrb1 antibody/product/Alomone Labs
Average 94 stars, based on 1 article reviews
Price from $9.99 to $1999.99

anti b1 bradykinin receptor bdkrb1 antibody - by Bioz Stars, 2022-08

94/100 stars

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B1R gene deletion prevents inflammation in the kidney during DOCA-salt-induced hypertension. DOCA-salt treatment for 3 weeks significantly increased mRNA expression of pro-inflammatory cytokines TNF, IL-6, IL-1β and chemokine MCP-1 in the kidney cortex of wild-type (WT) mice compared with sham mice. B1R knockout (B1RKO) mice did not show this DOCA-salt-induced increase in inflammatory markers. Gene expression measured by real time RT-PCR. Data was normalized to β-actin and presented as mean ± SEM ( n = 6, Two-way ANOVA, * P

Journal: Frontiers in Medicine

Article Title: Kinin B1 Receptor Mediates Renal Injury and Remodeling in Hypertension

doi: 10.3389/fmed.2021.780834

Figure Lengend Snippet: B1R gene deletion prevents inflammation in the kidney during DOCA-salt-induced hypertension. DOCA-salt treatment for 3 weeks significantly increased mRNA expression of pro-inflammatory cytokines TNF, IL-6, IL-1β and chemokine MCP-1 in the kidney cortex of wild-type (WT) mice compared with sham mice. B1R knockout (B1RKO) mice did not show this DOCA-salt-induced increase in inflammatory markers. Gene expression measured by real time RT-PCR. Data was normalized to β-actin and presented as mean ± SEM ( n = 6, Two-way ANOVA, * P

Article Snippet: Membranes were blocked with odyssey TBS blocking buffer (Licor) and immunoblotted overnight at 4°C with a specific antibody against B1R (#ABR-011, Alomone labs).

Techniques: Expressing, Mouse Assay, Knock-Out, Quantitative RT-PCR

B1R expression is increased and correlates with collagen expression in human kidney sections from hypertensive chronic kidney disease. Representative immunohistochemistry photomicrographs showing B1R specific immunoreactivity in kidney sections of healthy controls (A) , specifically in glomerulus (B) , tubules (C) and arteries (D) . The B1R specific immunoreactivity is increased in kidney sections from renal disease (E) , specifically in glomerulus (F) , tubules (G) and arteries (H) . Representative photomicrographs showing Masson Trichrome staining for collagen deposition in health control kidney section (I) with in glomerulus (J) , tubules (K) and arteries (L) . The collagen deposition was increased in kidney sections from the kidney disease subjects (M) , specifically in glomerulus (N) , tubules (O) and arteries (P) . Quantification data showing increased B1R (Q) and collagen (R) expression in kidney sections from renal disease subjects ( n = 5, Unpaired, 2-tailed t -test, * P

Journal: Frontiers in Medicine

Article Title: Kinin B1 Receptor Mediates Renal Injury and Remodeling in Hypertension

doi: 10.3389/fmed.2021.780834

Figure Lengend Snippet: B1R expression is increased and correlates with collagen expression in human kidney sections from hypertensive chronic kidney disease. Representative immunohistochemistry photomicrographs showing B1R specific immunoreactivity in kidney sections of healthy controls (A) , specifically in glomerulus (B) , tubules (C) and arteries (D) . The B1R specific immunoreactivity is increased in kidney sections from renal disease (E) , specifically in glomerulus (F) , tubules (G) and arteries (H) . Representative photomicrographs showing Masson Trichrome staining for collagen deposition in health control kidney section (I) with in glomerulus (J) , tubules (K) and arteries (L) . The collagen deposition was increased in kidney sections from the kidney disease subjects (M) , specifically in glomerulus (N) , tubules (O) and arteries (P) . Quantification data showing increased B1R (Q) and collagen (R) expression in kidney sections from renal disease subjects ( n = 5, Unpaired, 2-tailed t -test, * P

Article Snippet: Membranes were blocked with odyssey TBS blocking buffer (Licor) and immunoblotted overnight at 4°C with a specific antibody against B1R (#ABR-011, Alomone labs).

Techniques: Expressing, Immunohistochemistry, Staining

B1R blockade reduces reactive oxygen species (ROS) production in the kidney and HK-2 cells. (A) Representative photomicrographs showing dihydroethidium (DHE) staining in the kidney sections. (B) DHE staining quantified as relative fluorescence intensity showing increased superoxide generation in the kidneys of DOCA-salt treated hypertensive mice compared with sham treated wild-type (WT) mice. This increase in DHE staining was blunted in B1R deficient mice (B1RKO) mice with DOCA-salt treatment ( n = 6, Two-way ANOVA, * P

Journal: Frontiers in Medicine

Article Title: Kinin B1 Receptor Mediates Renal Injury and Remodeling in Hypertension

doi: 10.3389/fmed.2021.780834

Figure Lengend Snippet: B1R blockade reduces reactive oxygen species (ROS) production in the kidney and HK-2 cells. (A) Representative photomicrographs showing dihydroethidium (DHE) staining in the kidney sections. (B) DHE staining quantified as relative fluorescence intensity showing increased superoxide generation in the kidneys of DOCA-salt treated hypertensive mice compared with sham treated wild-type (WT) mice. This increase in DHE staining was blunted in B1R deficient mice (B1RKO) mice with DOCA-salt treatment ( n = 6, Two-way ANOVA, * P

Article Snippet: Membranes were blocked with odyssey TBS blocking buffer (Licor) and immunoblotted overnight at 4°C with a specific antibody against B1R (#ABR-011, Alomone labs).

Techniques: Staining, Fluorescence, Mouse Assay

B1R gene deletion prevents gene expression of renal fibrosis markers in DOCA-salt hypertension. Gene expression measured by real time RT-PCR showing significantly increased mRNA of renal fibrosis markers in the kidney cortex of DOCA-salt treated hypertensive mice compared with sham mice. B1R knockout mice did not show this DOCA-salt-induced increase in renal fibrosis markers. Data was normalized to β-actin and presented as mean ± SEM ( n = 6, Two-way ANOVA, * P

Journal: Frontiers in Medicine

Article Title: Kinin B1 Receptor Mediates Renal Injury and Remodeling in Hypertension

doi: 10.3389/fmed.2021.780834

Figure Lengend Snippet: B1R gene deletion prevents gene expression of renal fibrosis markers in DOCA-salt hypertension. Gene expression measured by real time RT-PCR showing significantly increased mRNA of renal fibrosis markers in the kidney cortex of DOCA-salt treated hypertensive mice compared with sham mice. B1R knockout mice did not show this DOCA-salt-induced increase in renal fibrosis markers. Data was normalized to β-actin and presented as mean ± SEM ( n = 6, Two-way ANOVA, * P

Article Snippet: Membranes were blocked with odyssey TBS blocking buffer (Licor) and immunoblotted overnight at 4°C with a specific antibody against B1R (#ABR-011, Alomone labs).

Techniques: Expressing, Quantitative RT-PCR, Mouse Assay, Knock-Out

B1R gene deletion abrogates DOCA-salt induced renal myofibroblast transition. (A) Representative immunofluorescence images showing expression of alpha-smooth muscle actin (α-SMA), fibronectin, and transforming growth factor-beta (TGF-β) in the kidney sections. Quantification data presented as mean fluorescence staining showing increased expression of α-SMA (B) , fibronectin (C) , and TGF-β (D) of DOCA-salt treated hypertensive mice compared with sham mice. These increases in fibrosis markers expression were blunted in B1R deficient mice with 3 weeks of DOCA-salt treatment ( n = 6, Two-way ANOVA, * P

Journal: Frontiers in Medicine

Article Title: Kinin B1 Receptor Mediates Renal Injury and Remodeling in Hypertension

doi: 10.3389/fmed.2021.780834

Figure Lengend Snippet: B1R gene deletion abrogates DOCA-salt induced renal myofibroblast transition. (A) Representative immunofluorescence images showing expression of alpha-smooth muscle actin (α-SMA), fibronectin, and transforming growth factor-beta (TGF-β) in the kidney sections. Quantification data presented as mean fluorescence staining showing increased expression of α-SMA (B) , fibronectin (C) , and TGF-β (D) of DOCA-salt treated hypertensive mice compared with sham mice. These increases in fibrosis markers expression were blunted in B1R deficient mice with 3 weeks of DOCA-salt treatment ( n = 6, Two-way ANOVA, * P

Article Snippet: Membranes were blocked with odyssey TBS blocking buffer (Licor) and immunoblotted overnight at 4°C with a specific antibody against B1R (#ABR-011, Alomone labs).

Techniques: Immunofluorescence, Expressing, Fluorescence, Staining, Mouse Assay

B1R gene deletion prevents kidney injury in DOCA-salt hypertension. (A) Representative immunofluorescence images showing expression of kidney injury molecule-1 (Kim-1) in the kidney sections. (B) Quantification data presented as mean fluorescence staining showing increased expression of Kim-1 in the kidneys of DOCA-salt treated hypertensive mice compared with sham treated wild-type (WT) mice. This increase in Kim-1 expression was blunted in B1R deficient mice (B1RKO) mice with DOCA-salt treatment ( n = 5, Two-way ANOVA, * P

Journal: Frontiers in Medicine

Article Title: Kinin B1 Receptor Mediates Renal Injury and Remodeling in Hypertension

doi: 10.3389/fmed.2021.780834

Figure Lengend Snippet: B1R gene deletion prevents kidney injury in DOCA-salt hypertension. (A) Representative immunofluorescence images showing expression of kidney injury molecule-1 (Kim-1) in the kidney sections. (B) Quantification data presented as mean fluorescence staining showing increased expression of Kim-1 in the kidneys of DOCA-salt treated hypertensive mice compared with sham treated wild-type (WT) mice. This increase in Kim-1 expression was blunted in B1R deficient mice (B1RKO) mice with DOCA-salt treatment ( n = 5, Two-way ANOVA, * P

Article Snippet: Membranes were blocked with odyssey TBS blocking buffer (Licor) and immunoblotted overnight at 4°C with a specific antibody against B1R (#ABR-011, Alomone labs).

Techniques: Immunofluorescence, Expressing, Fluorescence, Staining, Mouse Assay

B1R expression in human kidney proximal tubular epithelial (HK-2) cells. (A) Representative photomicrographs showing immunofluorescence of B1R in HK-2 cells treated vehicle or B1R specific agonist DAKD for 24 h. Control HK-2 cells without primary antibody (No 1°Ab control) incubation and with only secondary antibody incubation shows no staining. (B) The quantification of B1R immunoreactivity shows that the DAKD treatment of HK-2 cells for 24 h significantly upregulated the B1R expression ( n =7–8, Unpaired, 2-tailed t -test, * P

Journal: Frontiers in Medicine

Article Title: Kinin B1 Receptor Mediates Renal Injury and Remodeling in Hypertension

doi: 10.3389/fmed.2021.780834

Figure Lengend Snippet: B1R expression in human kidney proximal tubular epithelial (HK-2) cells. (A) Representative photomicrographs showing immunofluorescence of B1R in HK-2 cells treated vehicle or B1R specific agonist DAKD for 24 h. Control HK-2 cells without primary antibody (No 1°Ab control) incubation and with only secondary antibody incubation shows no staining. (B) The quantification of B1R immunoreactivity shows that the DAKD treatment of HK-2 cells for 24 h significantly upregulated the B1R expression ( n =7–8, Unpaired, 2-tailed t -test, * P

Article Snippet: Membranes were blocked with odyssey TBS blocking buffer (Licor) and immunoblotted overnight at 4°C with a specific antibody against B1R (#ABR-011, Alomone labs).

Techniques: Expressing, Immunofluorescence, Incubation, Staining

Elevated B1R expression in the kidneys of DOCA-salt hypertensive mice. (A) Gene expression measured by real time RT-PCR showing significantly increased mRNA in the kidneys at 7, 14, and 21 days of DOCA-salt treated hypertensive mice compared with sham mice ( n = 6, One-way ANOVA, * P

Journal: Frontiers in Medicine

Article Title: Kinin B1 Receptor Mediates Renal Injury and Remodeling in Hypertension

doi: 10.3389/fmed.2021.780834

Figure Lengend Snippet: Elevated B1R expression in the kidneys of DOCA-salt hypertensive mice. (A) Gene expression measured by real time RT-PCR showing significantly increased mRNA in the kidneys at 7, 14, and 21 days of DOCA-salt treated hypertensive mice compared with sham mice ( n = 6, One-way ANOVA, * P

Article Snippet: Membranes were blocked with odyssey TBS blocking buffer (Licor) and immunoblotted overnight at 4°C with a specific antibody against B1R (#ABR-011, Alomone labs).

Techniques: Expressing, Mouse Assay, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Activation of Kinin B1R Upregulates ADAM17 and Results in ACE2 Shedding in Neurons

doi: 10.3390/ijms22010145

Figure Lengend Snippet: Effect of B1R activation on ADAM17 activity in primary hypothalamic neurons. ( A ) Incubation of wild-type neurons with B1R-specific agonist Lys-Des-Arg 9 -Bradykinin (LDABK) significantly increased ADAM17 activity. Pre-treatment with R715, a B1R-specific antagonist, prevented this LDABK-mediated increase in ADAM17 activity. R715 treatment alone did not have any effect on ADAM17 activity. ( B ) Treatment with LDABK did not show any effect on ADAM17 activity in neurons with B1R deletion. In addition, the HOE 140, a kinin B2 receptor (B2R)-specific antagonist also did not show any effect on ADAM17 activity. The cultured primary neurons were pre-treated with a specific B1R antagonist (R715, 10 μM) or a specific B2R antagonist HOE 140 (HOE, 10 μM) for 1 h, followed by treatment with LDABK (300 nM) for 18 h ( n = 6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Article Snippet: Triple immunostaining was performed with specific validated antibodies for detection of ADAM17 (#ab13535, lot GR56873-25, Abcam, Cambridge, UK, 1:500 dilution) and B1R (#ABR-011, lot An-01, Alomone labs, Jerusalem, Israel, 1:200 dilution) or ACE2 ((#OASG00144, lot 1442701, Aviva systems biology, San Diego, CA, USA, 1:200 dilution) coupled with DAPI staining.

Techniques: Activation Assay, Activity Assay, Incubation, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Activation of Kinin B1R Upregulates ADAM17 and Results in ACE2 Shedding in Neurons

doi: 10.3390/ijms22010145

Figure Lengend Snippet: Effect of B1R activation on ACE2 activity in primary hypothalamic neurons. ( A ) Incubation of wild-type neurons with B1R-specific agonist LDABK significantly decreased ACE2 activity. R715, a B1R-specific antagonist pretreatment prevented this LDABK-mediated decrease in ACE2 activity. R715 treatment alone did not have any effect on ACE2 activity. ( B ) Treatment with LDABK did not show any effect on ACE2 activity in neurons with B1R deletion. In addition, the HOE 140, a B2R-specific antagonist also did not show any effect on ACE2 activity. The cultured primary neurons were pre-treated with a specific B1R antagonist (R715, 10 μM) or a specific B2R antagonist HOE 140 (HOE, 10 μM) for 1 h, followed by treatment with LDABK (300 nM) for 18 h ( n = 6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques: Activation Assay, Activity Assay, Incubation, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Activation of Kinin B1R Upregulates ADAM17 and Results in ACE2 Shedding in Neurons

doi: 10.3390/ijms22010145

Figure Lengend Snippet: Effect of glutamate on B1R activation in primary hypothalamic neurons. Representative triple immunostaining revealed that compared to vehicle ( A ) treatment, stimulation with glutamate (100 μM) ( B ) for 18 h induced the expression of kinin B1R and ADAM17 protein in primary hypothalamic neurons. This glutamate-induced increase in B1R and ADAM17 protein expression was prevented by pre-treatment with R715 ( C ). Treatment with glutamate (100 μM) for 4 h induced an increase in B1R mRNA in cultured neurons, measured by real time PCR. This increase in B1R mRNA was prevented by pre-treatment with R715 ( D ). Representative image of Western blot ( E ) and quantitative analysis ( F ) showing glutamate stimulation induced B1R protein expression which was prevented by pre-treatment with R715 in neurons ( n = 4–6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques: Activation Assay, Triple Immunostaining, Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Activation of Kinin B1R Upregulates ADAM17 and Results in ACE2 Shedding in Neurons

doi: 10.3390/ijms22010145

Figure Lengend Snippet: Kinin B1 receptor (B1R), A Disintegrin And Metalloprotease 17 (ADAM17), and angiotensin converting enzyme 2 (ACE2) expression in primary hypothalamic neurons. Representative photomicrographs showing immunofluorescence staining for neuron-specific marker microtubule associated protein 2 (MAP-2) (Green) in primary neurons from wild-type ( A ) and B1R knockout mice ( B ) cultured for 10 days. Primary neurons from wild-type mice pups showed immunopositivity for the presence of B1R ( C ), while B1R immunoreactivity was absent in neurons with B1R deletion ( D ). Representative triple immunostaining revealed that kinin B1R ( E ) and ADAM17 ( F ) are expressed in primary hypothalamic neurons expressing nuclear DAPI staining in blue ( G ), and merged image ( H ) shows the colocalization of B1R and ADAM17. Representative triple immunostaining showing ACE2 and ADAM17 expression and co-localization in wild-type neurons ( I – L ) and in B1R knockout neurons ( M – P ).

Techniques: Expressing, Immunofluorescence, Staining, Marker, Knock-Out, Mouse Assay, Cell Culture, Triple Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: Activation of Kinin B1R Upregulates ADAM17 and Results in ACE2 Shedding in Neurons

doi: 10.3390/ijms22010145

Figure Lengend Snippet: Activation of kinin B1R upregulates ADAM17-mediated ACE2 shedding in primary hypothalamic neurons. B1R activation by specific agonist results in upregulation of ADAM17 expression and activity which results in increased membrane-bound ACE2 shedding and reduced ACE2 activity in primary hypothalamic neurons. In addition, glutamate can induce B1R expression and B1R is involved in glutamate-mediated effects on ADAM17 activity and ACE2 shedding.

Techniques: Activation Assay, Expressing, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Activation of Kinin B1R Upregulates ADAM17 and Results in ACE2 Shedding in Neurons

doi: 10.3390/ijms22010145

Figure Lengend Snippet: Glutamate-induced ADAM17-mediated ACE2 shedding involves kinin B1R activation. Glutamate increased ADAM17 activity ( A ) and decreased ACE2 activity ( B ) in neurons, which was prevented by pre-treatment with R715. Treatment of B1R knockout neurons with glutamate did not show any effect on ADAM17 or ACE2 activity. ADAM17 and ACE2 assays were performed in primary hypothalamic neurons cultured from wild-type and B1R knockout neonates, treated with glutamate (Glut, 100 μM) or glutamate with 1 h of pre-treatment with R715 (a specific B1R inhibitor, 10 μM) for 18 h ( n = 6 independent cultures/group). Statistical significance: one-way ANOVA followed by Tukey’s multiple comparisons test. * p

Techniques: Activation Assay, Activity Assay, Knock-Out, Cell Culture

Endothelial expression of bradykinin receptor b1 as well as b2 in lung tissue. (A and E) Quantification data from immunofluorescence stainings. (B–D) Representative immunofluorescence images of bradykinin receptor b1 as well as ( F – H ) bradykinin receptor b2 staining in vessels of lung tissue. One-way ANOVA followed by Dunnett's post hoc test for significance vs. NaCl controls was used. Error bars indicate mean ± SD. *P

Journal: PLoS ONE

Article Title: C1 Esterase Inhibitor Reduces Lower Extremity Ischemia/Reperfusion Injury and Associated Lung Damage

doi: 10.1371/journal.pone.0072059

Figure Lengend Snippet: Endothelial expression of bradykinin receptor b1 as well as b2 in lung tissue. (A and E) Quantification data from immunofluorescence stainings. (B–D) Representative immunofluorescence images of bradykinin receptor b1 as well as ( F – H ) bradykinin receptor b2 staining in vessels of lung tissue. One-way ANOVA followed by Dunnett's post hoc test for significance vs. NaCl controls was used. Error bars indicate mean ± SD. *P

Article Snippet: Furthermore, we analyzed fibrin deposition (F0111; Dako, Baar, Switzerland), expression of heparan sulfate (HS; 370255, Amsbio, Abingdon, UK), bradykinin receptor b1 (ABR-011, Alomone Labs, Jerusalem, Israel), bradykinin receptor b2 (ABR-012, Alomone Labs) as well as VE-cadherin (sc-6458, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA).

Techniques: Expressing, Immunofluorescence, Staining