Application
Image of NPY Y2 receptor immunoreactivity in mouse dorsal root ganglion of wild type (A) and NPY Y2 receptor Knockout (B) animals. Note that unspecific staining is observed in thick, possibly myelinated fibers. Image courtesy of Dr. Tomas Hkfelt, KarolinsApplication
CCK regulates expression of Y2R in vagal afferent neurons. A, B, Expression of Y2R in rats fasted for 48 h. (A) is decreased in the mid-caudal regions of the nodose ganglion compared with rats fed ad libitum (B), although some neurons still express Y2R in fasted rats. C, After administration of CCK8s (10 nmol, i.p.) to fasted rats, there is a substantial increase of Y2R-positive neurons. D, For comparison, control samples show low abundance of Y2R 2 h after administration of vehicle. E, Refeeding of fasted rats for 2 h increases expression of Y2R in nodose neurons. F, In contrast, in rats treated with a CCK1R antagonist (lorglumide) 15 min before refeeding, the increase of Y2R after refeeding was inhibited. G–I, At higher power, dual staining with antibodies to Y2R (G, red) and glutamine synthetase (H, green) reveals that Y2R immunoreactivity is localized to satellite cells (open arrowheads) as well as neurons (filled arrows); I, overlay of panels G and H showing colocalization in satellite cells (yellow, filled arrowheads). J–L, In rats fed ad libitum, there is Y2R in both neurons (J, red, filled arrows) and satellite cells (K, green, open arrows; L, overlay with colocalization in satellite cells indicated by filled arrowheads). However, in fasted rats (M–O), there is decreased expression of Y2R in neuronal cell bodies (M, filled arrows) but not satellite cells (N, open arrows; O, overlay with colocalization in satellite cells indicated by filled arrowheads). P–T, Localization of Y2R in human nodose ganglia. P–R, Dual staining with antibodies to Y2R (P, red) and GFAP (Q, green) shows that Y2R is expressed in small and large neurons (red, filled arrows) as well as satellite cells (open arrows) (R, overlay with colocalization to satellite cells indicated by open arrowheads). S, Colocalization of Y2R (red) and CCK1R (green) in small and large neurons indicated by open arrows. T, Colocalization of Y2R (red) and VR1 (green) in small neurons (open arrows). U–W, Quantification of CCK-dependent changes in Y2R-positive neurons. U, Progressive decrease in the numbers of Y2R-immunoreactive cells in nodose ganglia after nutrient withdrawal. Exponential curve-fitting (OriginPro7.5) indicated 50% decrease in abundance of Y2R-immunoreactive neurons at 12.5 h (n = 4). V, Intraperitoneal administration of CCK8s (10 nmol) to rats fasted for 24 h increased the number of Y2R-immunoreactive neurons with a peak at 2 h (n = 4). W, Fasting for 24 h decreased the number of Y2R-immunoreactive neurons (***p| Product Name | NPY Y2 antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal NPY Y2 Receptor antiserumThis NPY Y2 antibody has been extensively characterized at the spinal level and is currently being characerized in brain.? The antibody has been tested in both rat and mouse and appears to work better on mose tissue, although positive staining is seen in rat.? For full characterizarion in dorsal root ganglion and spinal cord see: J Comp Neurol. 2005 Aug 29;489(3):328-48. |
| Clonality | Polyclonal |
|---|---|
| Host Species | Rabbit |
| Tested Applications | IHC |
| For IHC: Use at a dilution of 1:500 -1:1,000. Optimal dilutions/concentrations should be determined by the researcher. Antiserum was used on perfusion fixed tissue. Perfusion: 1) calcium-free Tyrode’s solution, 2) paraformaldehyde-picric acid fixative, and 3) 10% sucrose in PBS as a cryo-protectant. Desired tissues were dissected and stored overnight in 10% sucrose in PBS. Slide-mounted tissue sections were processed for indirect immunofluorescence. Slides were incubated with blocking buffer for 1 hour at room temperature. Primary antiserum was diluted with blocking buffer to the appropriate working concentration. Blocking buffer was removed and slides were incubated for 18-24 hours at 4oC with primary antiserum. Slides were rinsed 3 times and then incubated with secondary antibodies for 1 hour at room temperature. Slides were again rinsed 3 times and coverslipped. Staining was examined using fluorescence microscopy.: |
|
| Species Reactivity | MouseRat |
| Concentration | 1mg/ml |
| Target | NPY Y2 |
|---|
Application
Image of NPY Y2 receptor immunoreactivity in mouse dorsal root ganglion of wild type (A) and NPY Y2 receptor Knockout (B) animals. Note that unspecific staining is observed in thick, possibly myelinated fibers. Image courtesy of Dr. Tomas Hkfelt, Karolins
Application
CCK regulates expression of Y2R in vagal afferent neurons. A, B, Expression of Y2R in rats fasted for 48 h. (A) is decreased in the mid-caudal regions of the nodose ganglion compared with rats fed ad libitum (B), although some neurons still express Y2R in fasted rats. C, After administration of CCK8s (10 nmol, i.p.) to fasted rats, there is a substantial increase of Y2R-positive neurons. D, For comparison, control samples show low abundance of Y2R 2 h after administration of vehicle. E, Refeeding of fasted rats for 2 h increases expression of Y2R in nodose neurons. F, In contrast, in rats treated with a CCK1R antagonist (lorglumide) 15 min before refeeding, the increase of Y2R after refeeding was inhibited. G–I, At higher power, dual staining with antibodies to Y2R (G, red) and glutamine synthetase (H, green) reveals that Y2R immunoreactivity is localized to satellite cells (open arrowheads) as well as neurons (filled arrows); I, overlay of panels G and H showing colocalization in satellite cells (yellow, filled arrowheads). J–L, In rats fed ad libitum, there is Y2R in both neurons (J, red, filled arrows) and satellite cells (K, green, open arrows; L, overlay with colocalization in satellite cells indicated by filled arrowheads). However, in fasted rats (M–O), there is decreased expression of Y2R in neuronal cell bodies (M, filled arrows) but not satellite cells (N, open arrows; O, overlay with colocalization in satellite cells indicated by filled arrowheads). P–T, Localization of Y2R in human nodose ganglia. P–R, Dual staining with antibodies to Y2R (P, red) and GFAP (Q, green) shows that Y2R is expressed in small and large neurons (red, filled arrows) as well as satellite cells (open arrows) (R, overlay with colocalization to satellite cells indicated by open arrowheads). S, Colocalization of Y2R (red) and CCK1R (green) in small and large neurons indicated by open arrows. T, Colocalization of Y2R (red) and VR1 (green) in small neurons (open arrows). U–W, Quantification of CCK-dependent changes in Y2R-positive neurons. U, Progressive decrease in the numbers of Y2R-immunoreactive cells in nodose ganglia after nutrient withdrawal. Exponential curve-fitting (OriginPro7.5) indicated 50% decrease in abundance of Y2R-immunoreactive neurons at 12.5 h (n = 4). V, Intraperitoneal administration of CCK8s (10 nmol) to rats fasted for 24 h increased the number of Y2R-immunoreactive neurons with a peak at 2 h (n = 4). W, Fasting for 24 h decreased the number of Y2R-immunoreactive neurons (***p| Application Notes | For IHC: Use at a dilution of 1:500 -1:1,000. Optimal dilutions/concentrations should be determined by the researcher. Antiserum was used on perfusion fixed tissue. Perfusion: 1) calcium-free Tyrode’s solution, 2) paraformaldehyde-picric acid fixative, and 3) 10% sucrose in PBS as a cryo-protectant. Desired tissues were dissected and stored overnight in 10% sucrose in PBS. Slide-mounted tissue sections were processed for indirect immunofluorescence. Slides were incubated with blocking buffer for 1 hour at room temperature. Primary antiserum was diluted with blocking buffer to the appropriate working concentration. Blocking buffer was removed and slides were incubated for 18-24 hours at 4oC with primary antiserum. Slides were rinsed 3 times and then incubated with secondary antibodies for 1 hour at room temperature. Slides were again rinsed 3 times and coverslipped. Staining was examined using fluorescence microscopy.: |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Keep as concentrated solution. Aliquot and store at -20ºC or below. Avoid freeze-thaw cycles. |
| Storage Buffer | Phosphate-buffered saline containing 0.05% sodium azide |
