polyclonal rabbit antibody  (Alomone Labs)

Journal: Annals of neurology

Article Title: Encephalitis and antibodies to DPPX, a subunit of Kv4.2 potassium channels

doi: 10.1002/ana.23756

Figure Lengend Snippet: Immunoprecipitation of DPPX In cultures of dissociated rat hippocampal neurons, patients’ antibodies showed intense reactivity with the neuronal cell surface (A), bar = 10 μm. Immunoprecipitation of the antigen with serum of the index case is shown in B, where the precipitated proteins were run in a gel and subsequently stained with EZblue. Note that patient’s antibodies precipitated a protein (band close to 102 kDa in lane P), which was excised from the gel and analyzed by mass spectrometry, demonstrating sequences of DPPX. Lane N is the precipitate obtained from control serum. Immunoblot of these proteins with a rabbit polyclonal antibody against DPPX (1:1000, developed by BR) confirmed that the band corresponded to DPPX (C).

Article Snippet: Similar studies comparing the serum of a healthy individual and the rabbit polyclonal antibody are shown in M and N, and the merged images in O.

Techniques: Immunoprecipitation, Staining, Mass Spectrometry

Journal: Annals of neurology

Article Title: Encephalitis and antibodies to DPPX, a subunit of Kv4.2 potassium channels

doi: 10.1002/ana.23756

Figure Lengend Snippet: Expression of DPPX in myenteric plexus Transverse section of small bowel of rat showing the longitudinal muscular layer (LM), circular muscular layer (CM), submucosal layer (SM), and glans (G). The myenteric plexus (Plex) is revealed as clusters of large neurons between the two muscular layers. In the 3 panels (A–C) the nuclei of the neurons (red) was labeled with anti-Hu (a highly specific neuronal marker). Panel A, shows in green the DPPX immunostaining using a rabbit polyclonal antibody (1:1000, developed by BD); panel B shows the DPPX reactivity of serum from one of the patients with encephalitis, and panel C shows the lack of reactivity of serum from a healthy subject. Note that DPPX is predominantly expressed in the cytoplasm-membrane of the large clustered neurons of the myenteric plexus, and is also detected in a fine longitudinal pattern in CM and SM where the submucosal plexus is located. Bar = 20μm.

Article Snippet: Similar studies comparing the serum of a healthy individual and the rabbit polyclonal antibody are shown in M and N, and the merged images in O.

Techniques: Expressing, Labeling, Marker, Immunostaining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Increased rhythmicity in hypertensive arterial smooth muscle is linked to transient receptor potential canonical channels

doi: 10.1111/j.1582-4934.2009.00890.x

Figure Lengend Snippet: Effect of candesartan or telmisartan but not amlodipine on TRPC expression in mesenteric arterioles. Long-term administration of angiotensin AT1 receptor antagonist telmisartan or candesartan, but not of calcium channel blocker amlodipine reduces TRPC1, TRPC3 and TRPC5 channel protein expression in vivo. The angiotensin AT1 receptor antagonist telmisartan (5 mg/kg per day) or candesartan (4 mg/kg per day), calcium channel blocker amlodipine (10 mg/kg per day), or placebo were administered to SHR by gavage for 16 weeks. Representative immunoblottings of TRPC channel protein expressions in mesenteric arterioles from treated SHR (Fig. 7A) and summary data are shown (Fig. 7B). * P

Article Snippet: The membranes were incubated with primary rabbit antibodies against TRPC1, TRPC3, TRPC4, TRPC5 and TRPC6 (1:200; Alomone Labs), followed by incubation with goat anti-rabbit horseradish peroxidase-conjugated secondary antibody.

Techniques: Expressing, In Vivo

Journal: Journal of Cellular and Molecular Medicine

Article Title: Increased rhythmicity in hypertensive arterial smooth muscle is linked to transient receptor potential canonical channels

doi: 10.1111/j.1582-4934.2009.00890.x

Figure Lengend Snippet: Inhibition of norepinephrine-induced vasomotion in mesenteric arterioles from SHR by specific anti-TRPC antibodies Representative tracings of norepinephrine-induced vasomotion in mesenteric arterioles from SHR under control conditions (A), in the presence of anti-TRPC1 antibodies (B), anti-TRPC3 antibodies (C), anti-TRPC3 antibodies with antigenic peptide (D), anti-TRPC5 antibodies (E), anti-TRPC1 plus anti-TRPC3 antibodies (F) or in the presence of anti-β-actin antibodies (G). Representative tracings of norepinephrine-induced vasomotion in mesenteric arterioles from SHR under control conditions after short-term exposure to hypotonic 0.45% NaCl without antibody (H), or after short-term exposure to hypotonic 0.45% NaCl plus TRPC3 antibody (I). Summary data (J) show that inhibition of TRPC channels by specific anti-TRPC antibodies significantly attenuates norepinephrine-induced vasomotion in SHR. Data are mean ± S.E.M. of n = 6 independent experiments. * P

Article Snippet: The membranes were incubated with primary rabbit antibodies against TRPC1, TRPC3, TRPC4, TRPC5 and TRPC6 (1:200; Alomone Labs), followed by incubation with goat anti-rabbit horseradish peroxidase-conjugated secondary antibody.

Techniques: Inhibition

Journal: Journal of Cellular and Molecular Medicine

Article Title: Increased rhythmicity in hypertensive arterial smooth muscle is linked to transient receptor potential canonical channels

doi: 10.1111/j.1582-4934.2009.00890.x

Figure Lengend Snippet: Expression of TRPC1, TRPC3 and TRPC5 channels in mesenteric arterioles from SHR. Representative immunoblottings (A) and summary data (B) of TRPC1, TRPC3, TRPC4, TRPC5,TRPC6 channels and TRPC3 antibody with its respective antigenic peptide the protein expression in mesenteric arterioles from WKY (open bars) and SHR (filled bars). Data are mean ± S.E.M. of n = 4 independent experiments. * P

Article Snippet: The membranes were incubated with primary rabbit antibodies against TRPC1, TRPC3, TRPC4, TRPC5 and TRPC6 (1:200; Alomone Labs), followed by incubation with goat anti-rabbit horseradish peroxidase-conjugated secondary antibody.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Increased rhythmicity in hypertensive arterial smooth muscle is linked to transient receptor potential canonical channels

doi: 10.1111/j.1582-4934.2009.00890.x

Figure Lengend Snippet: Inhibition of norepinephrine-induced calcium increase in mesenteric arterioles from SHR by specific anti-TRPC antibodies. Representative tracings of norepinephrine-induced calcium increase in mesenteric arterioles from SHR under control conditions and in the presence of anti-TRPC1 antibodies (A), anti-TRPC3 antibodies (B), anti-TRPC5 antibodies (C), anti-TRPC1 plus anti-TRPC3 plus anti-TRPC5 antibodies (D), anti-TRPC3 antibodies plus TRPC3 antigenic peptide (E) or anti-β-actin antibodies (F). Summary data (G) shows the effects of specific anti-TRPC antibodies on norepinephrine-induced calcium influx in mesenteric arterioles from SHR. Data are mean ± S.E.M. of n = 8 independent experiments. * P

Article Snippet: The membranes were incubated with primary rabbit antibodies against TRPC1, TRPC3, TRPC4, TRPC5 and TRPC6 (1:200; Alomone Labs), followed by incubation with goat anti-rabbit horseradish peroxidase-conjugated secondary antibody.

Techniques: Inhibition

Journal: Nucleic Acids Research

Article Title: A single anti-microRNA antisense oligodeoxyribonucleotide (AMO) targeting multiple microRNAs offers an improved approach for microRNA interference

doi: 10.1093/nar/gkn1053

Figure Lengend Snippet: MTg-AMO as an improved AMO approach for miRNA target finding. ( A and B ) Western blot analysis of the protein levels of TGFBI, APC BCL2L11, HCN2 and Cav1.2. For MTg-AMO 21/155/17 , human breast cancer MCF-7 cells were used and for MTg-AMO 1/133 , cultured neonatal rat ventricular myocytes were used. NC and NC MTg-AMO: negative control MTg-AMO. The relevant miRNA for each target mRNA is specified above the name of the target mRNA in western blot images. * P

Article Snippet: The sample was incubated overnight at 4°C with primary antibodies for TGFBI, APC and BCL2L11 (Santa Cruz), and for HCN2 and Cav1.2 (Alomone Labs).

Techniques: Western Blot, Cell Culture, Negative Control

Journal: Digestive Diseases and Sciences

Article Title: Involvement of EP2 and EP4 Receptors in Eosinophilic Esophagitis: A Pilot Study

doi: 10.1007/s10620-019-05623-5

Figure Lengend Snippet: Chemotaxis and adhesion assays with human blood eosinophils. Eosinophils from healthy donors were incubated with 100 nM of EP4 agonist ONO-AE1-329 or with 100 nM of the EP4 antagonist ONO-AE3-208 ( a ), with 100 and 300 nM of EP2 agonist butaprost ( b ), or with 100 nM of DP1 agonist BW245C and 1 µM of DP1 antagonist MK0524 ( c ), and chemotaxis was assessed using esophageal epithelial cell supernatant for chemoattraction. n = 6–12 (for a , and b ) and n = 3 ( c ); values are mean ± SEM; one-way ANOVA; Tukey’s post hoc test. *** p values

Article Snippet: For flow cytometric visualization of the EP receptors, cells were incubated with rabbit polyclonal antibodies against EP1, EP2, and EP3 (20 μg/ml; Alomone Labs, Jerusalem, Israel) and mouse polyclonal antibody against EP4 (20 μg/ml; Alomone Labs, Jerusalem, Israel), or normal rabbit or mouse IgG (20 μg/ml; Santa Cruz Biotechnology, Dallas, TX, USA) for 30 min at 4 °C.

Techniques: Chemotaxis Assay, Incubation

Journal: Digestive Diseases and Sciences

Article Title: Involvement of EP2 and EP4 Receptors in Eosinophilic Esophagitis: A Pilot Study

doi: 10.1007/s10620-019-05623-5

Figure Lengend Snippet: Immunohistochemistry shows strong DP1 staining in eosinophils ( a , arrows ), while epithelial cells in esophageal mucosal biopsies are practically devoid of staining. EP4 ( b ) and EP2 ( c ) immunostaining is prominent in eosinophils ( arrows ) but also visible in epithelial cells ( arrowheads ). 3-3′-Diaminobenzidine (DAB) was used as visualization substrate for DP1 and EP4 staining (brown precipitates), while for EP2, ImmPact™ NovaRED substrate was used (red precipitate). Calibration bar: 50 µm

Article Snippet: For flow cytometric visualization of the EP receptors, cells were incubated with rabbit polyclonal antibodies against EP1, EP2, and EP3 (20 μg/ml; Alomone Labs, Jerusalem, Israel) and mouse polyclonal antibody against EP4 (20 μg/ml; Alomone Labs, Jerusalem, Israel), or normal rabbit or mouse IgG (20 μg/ml; Santa Cruz Biotechnology, Dallas, TX, USA) for 30 min at 4 °C.

Techniques: Immunohistochemistry, Staining, Immunostaining

Journal: Digestive Diseases and Sciences

Article Title: Involvement of EP2 and EP4 Receptors in Eosinophilic Esophagitis: A Pilot Study

doi: 10.1007/s10620-019-05623-5

Figure Lengend Snippet: Flow cytometric evaluation of EP (EP1–4) and DP (DP1, DP2) receptor expression in human blood eosinophils. Changes in receptor expression were recorded as mean fluorescence intensity and data are expressed as fold increase in fluorescence over isotype control. Expression levels of EP2 and EP4 are significantly lower in patients with eosinophilic esophagitis (EoE) compared to healthy control subjects (control). Expression levels of EP1, EP3, DP1, and DP2 do not differ between EoE and controls. Values are mean ± SEM; Student’s t test; n = 6–7. * p

Article Snippet: For flow cytometric visualization of the EP receptors, cells were incubated with rabbit polyclonal antibodies against EP1, EP2, and EP3 (20 μg/ml; Alomone Labs, Jerusalem, Israel) and mouse polyclonal antibody against EP4 (20 μg/ml; Alomone Labs, Jerusalem, Israel), or normal rabbit or mouse IgG (20 μg/ml; Santa Cruz Biotechnology, Dallas, TX, USA) for 30 min at 4 °C.

Techniques: Expressing, Fluorescence

Journal: Digestive Diseases and Sciences

Article Title: Involvement of EP2 and EP4 Receptors in Eosinophilic Esophagitis: A Pilot Study

doi: 10.1007/s10620-019-05623-5

Figure Lengend Snippet: ECIS ® cell impedance assays were performed in esophageal epithelial cells with 10 and 300 nM of EP4 agonist ONO-AE1-329 ( a ) and EP2 agonist butaprost ( b ) (or medium as control). The decrease in resistance correlates with a loss in the monolayer’s integrity indicative of cell barrier decrease. n = 5–9; values are mean ± SEM; one-way-ANOVA; Tukey’s post hoc test; * p

Article Snippet: For flow cytometric visualization of the EP receptors, cells were incubated with rabbit polyclonal antibodies against EP1, EP2, and EP3 (20 μg/ml; Alomone Labs, Jerusalem, Israel) and mouse polyclonal antibody against EP4 (20 μg/ml; Alomone Labs, Jerusalem, Israel), or normal rabbit or mouse IgG (20 μg/ml; Santa Cruz Biotechnology, Dallas, TX, USA) for 30 min at 4 °C.

Techniques: Electric Cell-substrate Impedance Sensing