Journal: PLoS ONE

Article Title: Rat Hypocretin/Orexin Neurons Are Maintained in a Depolarized State by TRPC Channels

doi: 10.1371/journal.pone.0015673

Figure Lengend Snippet: (A–C) In voltage-clamp configuration, intracellular application of either a TRPC1 (A) or a TRPC3 (B) or a TRPC4 (C) antibody (AbTRPC1, AbTRPC3 and AbTRPC4 respectively, all from Alomone Labs) has no effect on the current needed to clamp the cell at −50 mV. (D–E) Intracellular application of a TRPC5 antibody (Alomone Labs), in contrast, induces an outward current (D) that is blocked by co-application of a TRPC5 antigen (E). (F–G) In current-clamp configuration, intracellular application of a TRPC5 antibody alone (from either Alomone Labs (F) or NeuroMab (inset in F) induces an important membrane hyperpolarization and cessation of firing. In contrast, intracellular co-application of the TRPC5 antibody (Alomone Labs) together with its appropriate antigen (AgTRPC5) has no effect (G, to compare with F). (H) Voltage-clamp ramps in absence (Ctrl) and presence of a TRPC5 antibody (AbTRPC5, Alomone Labs). In the inset, subtraction of voltage-clamp ramps (Ctrl - AbTRPC5) suggests the presence of a voltage-dependent TRPC5 current.

Article Snippet: When used, the TRPC5 control antigen (Alomone Labs) was added (at 12 or 18 ng/µl) to the TRPC5 antibody in the pipette.

Techniques:

Journal: PLoS ONE

Article Title: Rat Hypocretin/Orexin Neurons Are Maintained in a Depolarized State by TRPC Channels

doi: 10.1371/journal.pone.0015673

Figure Lengend Snippet: (A) Transient (2 min) bath-application of 100 µM flufenamic acid (FFA), a non specific blocker of cationic currents, produces a strong membrane hyperpolarization and cessation of firing; inset: in voltage-clamp this hyperpolarization corresponds to an outward current. (B) Voltage-clamp ramps in absence (Ctrl) and presence (FFA) of flufenamic acid. The subtraction of voltage-clamp ramps shown in the inset suggests the presence of a voltage-dependent cationic current. (C–D) In hcrt/orx neurons loaded with a TRPC5 antibody, bath-application of FFA had only a small additional effect either in current-clamp (C) or in voltage-clamp (D).

Article Snippet: When used, the TRPC5 control antigen (Alomone Labs) was added (at 12 or 18 ng/µl) to the TRPC5 antibody in the pipette.

Techniques:

Journal: PLoS ONE

Article Title: Rat Hypocretin/Orexin Neurons Are Maintained in a Depolarized State by TRPC Channels

doi: 10.1371/journal.pone.0015673

Figure Lengend Snippet: (A–B) Photomicrographs from a dual-immunostained section combining an hcrt/orx antibody from Santa Cruz (A) and a TRPC5 antibody from Alomone Labs (B). Examples of double labelled hcrt/orx-TRPC5 cells are indicated by white arrowheads and one such cell by a double circle (enlarged in the corresponding small panels to the right). Some hcrt/orx cells do not express TRPC5, as shown by an example indicated with a single circle. It is noteworthy that many hcrt/orx-negative cells also expressed TRPC5, as shown by an example indicated with a downward arrow (in panels A and B). (C–D) Same as in A and B with an hcrt/orx antibody from Phoenix Pharmaceuticals (C) and a TRPC5 antibody from NeuroMab (D). (Scale bars: 20 µm).

Article Snippet: When used, the TRPC5 control antigen (Alomone Labs) was added (at 12 or 18 ng/µl) to the TRPC5 antibody in the pipette.

Techniques:

Journal: PLoS ONE

Article Title: Effects of Melanocortin 1 Receptor Agonists in Experimental Nephropathies

doi: 10.1371/journal.pone.0087816

Figure Lengend Snippet: (A) Five different doses of adriamycin, in the range of 5–10 mg/kg, were intravenously injected and the level of albuminuria was measured after 7 days. The urinary albumin-to-creatinine ratio (UACR) increased with increasing levels of adriamycin. (B) Albuminuria at day 7. Treatment with the MC1R agonist BMS-470539 did not reduce the level of albuminuria for the 8 mg/kg adriamycin dose, on the contrary it slightly increased albuminuria for the 10 mg/kg dose (n = 17–19, p<0.05). The unspecific melanocortin receptor agonist α-MSH did not have any significant effect (n = 9, n.s.) compared with untreated adriamycin mice. Results are presented as mean ± SEM.

Article Snippet: An MC1R control peptide antigen (Alomone Labs, Ltd., Israel) was separately added at a 1∶1 weight ratio to the MC1R antibody solution before incubation to confirm antibody specificity.

Techniques: Injection

Journal: PLoS ONE

Article Title: Effects of Melanocortin 1 Receptor Agonists in Experimental Nephropathies

doi: 10.1371/journal.pone.0087816

Figure Lengend Snippet: Gene expression of MC1R in human, rat and mouse kidney tissue.

Article Snippet: An MC1R control peptide antigen (Alomone Labs, Ltd., Israel) was separately added at a 1∶1 weight ratio to the MC1R antibody solution before incubation to confirm antibody specificity.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Effects of Melanocortin 1 Receptor Agonists in Experimental Nephropathies

doi: 10.1371/journal.pone.0087816

Figure Lengend Snippet: Western blotting was performed in order to detect MC1R protein expression in mouse and human tissue. Left panel: incubation with a rabbit polyclonal MC1R antibody (Alomone Labs). Right panel: incubation with MC1R antibody and control blocking peptide antigen (BP) in order to show antibody specificity. A) mouse whole glomerular lysate, B) cultured wild type mouse podocytes and C) a human malignant melanoma cell line A375. MC1R protein expression was detected at the expected size of 37 kDa (left panel).

Article Snippet: An MC1R control peptide antigen (Alomone Labs, Ltd., Israel) was separately added at a 1∶1 weight ratio to the MC1R antibody solution before incubation to confirm antibody specificity.

Techniques: Western Blot, Expressing, Incubation, Blocking Assay, Cell Culture