rabbit anti b2r  (Alomone Labs)

Journal: The Journal of Neuroscience

Article Title: The NGFR100W Mutation Specifically Impairs Nociception without Affecting Cognitive Performance in a Mouse Model of Hereditary Sensory and Autonomic Neuropathy Type V

doi: 10.1523/JNEUROSCI.0688-19.2019

Figure Lengend Snippet: Analysis of DRG markers, sympathetic neurons and innervation, and DRG transcriptome of NGF R100W/wt mice. A , B , Normal expression of NF200, IB4, and CGRP in DRG neurons in both juveniles and adults ( A ) and in P5 mice ( B ). Scale bars, 100 μm. C , No significant difference in SCG cell numbers from NGF R100W/wt and NGF wt/wt mice; Student's two-tailed t test: t = 0.482, p = 0.650; NGF wt/wt , n = 4; NGF R100W/wt , n = 3. Scale bar, 100 μm. D , Normal sympathetic innervation of target internal organs in NGF R100W/wt mice with respect to NGF wt/wt controls; heart, Student's two-tailed t test: t = −0.268, p = 0.796; NGF wt/wt , n = 6; NGF R100W/wt , n = 4; stomach, Student's two-tailed t test: t = 0.216, p = 0.835; NGF wt/wt , n = 5; NGF R100W/wt , n = 4; kidney, Student's two-tailed t test: t = −0.002, p = 0.999; NGF wt/wt , n = 6; NGF R100W/wt , n = 4; spleen, Student's two-tailed t test: t = −0.165, p = 0.874; NGF wt/wt , n = 5; NGF R100W/wt , n = 3. Scale bars, 1 mm. E , Reduced expression of B2R and TRPV1 in DRG neurons of adult mice. Scale bar, 50 μm. B2R, Student's two-tailed t test: t = 6.219, ** p = 0.003; NGF wt/wt , n = 3; NGF R100W/wt , n = 3. TRPV1, Student's two-tailed t test: t = 12.455, *** p

Article Snippet: The antibodies and dilutions used were as follows: 1:500 mouse anti-NF200 (Sigma-Aldrich); 1:200 mouse anti-calcitonin gene-related peptide (CGRP; Rockland); 1:100 Invitrogen isolectin GS-B4-biotin conjugate (Thermo Fisher Scientific); 1:200 rabbit anti-B2R (Alomone Labs); 1:300 mouse anti-TRPV1 (Millipore); 1:200 Dako rabbit anti-PGP 9.5 (Agilent Technologies); and 1:300 rabbit anti-NGF M20 (Santa Cruz Biotechnology).

Techniques: Mouse Assay, Expressing, Two Tailed Test

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

Journal: Cells

Article Title: Involvement of Bradykinin Receptor 2 in Nerve Growth Factor Neuroprotective Activity

doi: 10.3390/cells9122651

Figure Lengend Snippet: Expression of B2R in mixed cortical cultures. Representative immunofluorescence images of cultured CNs stained with antibodies for B2R (red), neurons (MAP2, green), astrocytes (GFAP, green), or microglial cells (Iba1, green) and nuclei (Hoechts, blue) in control conditions (CTR) and after NGF treatment (+NGF). B2R immunoreactivity significantly increased in microglial cell body after NGF treatment. Scale bar: 15 µm.

Article Snippet: Protein concentration was measured using a Biorad DC protein assay kit (Bio-Rad, Hercules, CA, USA), and equivalent amounts of protein (30 μg) were separated on 4–12% Bis-Tris NuPage precast gels (Invitrogen), blocked with 5% milk in TBS-Tween for 1 h and then incubated overnight with rabbit anti-B2R antibody (Alomone labs 1:1000) or mouse anti-β-actin (Sigma Aldrich 1:10,000).

Techniques: Expressing, Immunofluorescence, Cell Culture, Staining

Journal: Cells

Article Title: Involvement of Bradykinin Receptor 2 in Nerve Growth Factor Neuroprotective Activity

doi: 10.3390/cells9122651

Figure Lengend Snippet: mRNA expression profiles of BK and its receptors genes after NGF treatment, deprivation, and rescue. Transcript levels of genes encoding BK ( Kng1 ; NM_012696), Bradykinin receptor 2 ( Bdkrb2 ; NM_001270713), and Bradykinin receptor 1 ( Bdkrb1 ; NM_030851) in cortical neurons (CNs) following 48 h NGF treatment (+NGF), induction of apoptosis by 6 h NGF deprivation (−NGF 6 h), and rescue by 6 h NGF replacement (+NGF 6 h). Data represent means (±S.E.M.) from four replicates. Statistically significant differences were calculated by one-way analysis of variance (ANOVA) followed by Bonferroni’s test for multiple comparison (* p

Article Snippet: Protein concentration was measured using a Biorad DC protein assay kit (Bio-Rad, Hercules, CA, USA), and equivalent amounts of protein (30 μg) were separated on 4–12% Bis-Tris NuPage precast gels (Invitrogen), blocked with 5% milk in TBS-Tween for 1 h and then incubated overnight with rabbit anti-B2R antibody (Alomone labs 1:1000) or mouse anti-β-actin (Sigma Aldrich 1:10,000).

Techniques: Expressing

Journal: Cells

Article Title: Involvement of Bradykinin Receptor 2 in Nerve Growth Factor Neuroprotective Activity

doi: 10.3390/cells9122651

Figure Lengend Snippet: Expression of B2R after NGF treatment and deprivation in CNs. At 10 DIV, CNs cultured in Neurobasal + 1% B27 (CTR) were treated for 48 h with NGF (100 ng/mL, +NGF) before being deprived of NGF by anti-NGF antibody treatment and incubated for 24 h (−NGF). B2R protein expression was measured by Western blot analysis. The immunoreactive signals at 45 kDa were quantified and normalized against β-actin and expressed as a percentage of the control (CTR). Data represent means (±S.E.M.) from four independent experiments run in duplicate. Statistically significant differences were calculated by one-way analysis of variance (ANOVA) followed by Bonferroni’s test for multiple comparisons (** p

Article Snippet: Protein concentration was measured using a Biorad DC protein assay kit (Bio-Rad, Hercules, CA, USA), and equivalent amounts of protein (30 μg) were separated on 4–12% Bis-Tris NuPage precast gels (Invitrogen), blocked with 5% milk in TBS-Tween for 1 h and then incubated overnight with rabbit anti-B2R antibody (Alomone labs 1:1000) or mouse anti-β-actin (Sigma Aldrich 1:10,000).

Techniques: Expressing, Cell Culture, Incubation, Western Blot

Journal: Cells

Article Title: Involvement of Bradykinin Receptor 2 in Nerve Growth Factor Neuroprotective Activity

doi: 10.3390/cells9122651

Figure Lengend Snippet: Overexpression of B2R by NGF treatment in 5xFAD brain extracts. Western blot analysis of B2R protein expression (panel ( a ) bands at 45 kDa; panel ( b ) bands at 70kDa) in brain extracts from wild type mice (CTR) and 5xFAD mice intranasally treated with PBS or NGF normalized against rpS6 and expressed as a percentage of the control (CTR). Data represent means (±S.E.M.) from four independent samples. Statistically significant differences were calculated by one-way analysis of variance (ANOVA) followed by Bonferroni’s test for multiple comparisons. (*** p

Article Snippet: Protein concentration was measured using a Biorad DC protein assay kit (Bio-Rad, Hercules, CA, USA), and equivalent amounts of protein (30 μg) were separated on 4–12% Bis-Tris NuPage precast gels (Invitrogen), blocked with 5% milk in TBS-Tween for 1 h and then incubated overnight with rabbit anti-B2R antibody (Alomone labs 1:1000) or mouse anti-β-actin (Sigma Aldrich 1:10,000).

Techniques: Over Expression, Western Blot, Expressing, Mouse Assay

Journal: Cells

Article Title: Involvement of Bradykinin Receptor 2 in Nerve Growth Factor Neuroprotective Activity

doi: 10.3390/cells9122651

Figure Lengend Snippet: Expression of bradykinin receptor 2 (B2R) after NGF treatment in cortical microglial cells. ( a ) Western blot analysis of B2R protein expression in enriched microglial cells cultures treated for 48 h with NGF 100 ng/mL (+NGF), normalized against β-actin, and expressed as a percentage of the control (CTR). Data represent means (±S.E.M.) from four independent experiments run in duplicate. Statistically significant differences were calculated by one-way analysis of variance (ANOVA) followed by Bonferroni’s test for multiple comparisons (* p

Article Snippet: Protein concentration was measured using a Biorad DC protein assay kit (Bio-Rad, Hercules, CA, USA), and equivalent amounts of protein (30 μg) were separated on 4–12% Bis-Tris NuPage precast gels (Invitrogen), blocked with 5% milk in TBS-Tween for 1 h and then incubated overnight with rabbit anti-B2R antibody (Alomone labs 1:1000) or mouse anti-β-actin (Sigma Aldrich 1:10,000).

Techniques: Expressing, Western Blot

Journal: Cells

Article Title: Involvement of Bradykinin Receptor 2 in Nerve Growth Factor Neuroprotective Activity

doi: 10.3390/cells9122651

Figure Lengend Snippet: Effect of B2R antagonism on CN viability. At 10 DIV, CNs cultured in Neurobasal + 1% B27 (CTR) were treated for 48 h with NGF 100 ng/mL (+NGF) and afterwards deprived of NGF for 24 h by washing (−NGF) or rescued by 24 h NGF replacement (+NGF). Neuronal viability was determined by counting intact nuclei and expressed considering 100 as the number of viable neurons in CTR conditions. ( a ) Treatment with HOE140 (1 µM), a selective B2R antagonist, induced a considerable inhibitory effect on the neuroprotection promoted by NGF replacement (HOE140 +NGF). ( b ) In the same conditions, HOE140 significantly antagonized the neuroprotective activity exerted by RPM-7, a selective B2R agonist (RPM-7 (100 nM). Data represent means (±S.E.M.) from four duplicate experiments. Statistically significant differences were calculated by one-way analysis of variance (ANOVA) followed by Bonferroni’s test (** p

Article Snippet: Protein concentration was measured using a Biorad DC protein assay kit (Bio-Rad, Hercules, CA, USA), and equivalent amounts of protein (30 μg) were separated on 4–12% Bis-Tris NuPage precast gels (Invitrogen), blocked with 5% milk in TBS-Tween for 1 h and then incubated overnight with rabbit anti-B2R antibody (Alomone labs 1:1000) or mouse anti-β-actin (Sigma Aldrich 1:10,000).

Techniques: Cell Culture, Activity Assay