Immunofluorescence analysis of NMDA Receptor 2B was done on 70% confluent log phase SHSY5Y cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with NMDA Receptor 2B Rabbit Polyclonal Antibody (PA3105) at 1ug/ml in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing membranous localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phosphopeptide corresponding to residues C G(1328) R F M D G S P Y(p) H(1337) of rat NMDA receptor 2B.|
|Storage buffer||whole serum|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunohistochemistry (IHC)||See 1 publications below|
PA3-105 detects N-methyl-D-aspartate (NMDA) receptor type 2B.
PA3-105 has been used successfully in Western blot, ELISA, immunoprecipitation, immunohistochemistry, and immunocytochemistry procedures. In Western blot analysis of rat brain synaptic membranes this antibody detects a ~180 kDa protein representing NMDA receptor type 2B.
The PA3-105 immunogen is a synthetic phosphopeptide corresponding to residues (C) G(1328) R F M D G S P Y(p) H(1337) of rat NMDA receptor 2B.
The ion channels activated by glutamate that are sensitive to N-methyl-D-aspartate (NMDA) are designated NMDA receptors (NMDAR). The NMDAR plays an essential role in memory, neuronal development and it has also been implicated in several disorders of the central nervous system including Alzheimer and quote;s, epilepsy and ischemic neuronal cell death (Grosshans et al., 2002; Wenthold et al., 2003; Carroll and Zukin, 2002). The NMDA receptor is also one of the principal molecular targets for alcohol in the CNS (Lovinger et al., 1989; Alvestad et al., 2003; Snell et al., 1996). The rat NMDAR1 (NR1) was the first subunit of the NMDAR to be clonedand it can form NMDA activated channels when expressed in Xenopus oocytes but the currents in such channels are much smaller than those seen in situ. Channels with more physiological characteristics are produced when the NR1 subunit is combined with one or more of the NMDAR2 (NR2 A-D) subunits. Overexpression of the NR2B-subunit of the NMDA receptor has been associated with increases in learning and memory while aged, memory impaired animals have deficiencies in NR2B expression (Clayton et al., 2002a; Clayton et al., 2002b). The NMDAR is also potentiated by protein phosphorylation (Lu et al., 1999).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Hijacking the neuronal NMDAR signaling circuit to promote tumor growth and invasion.
PA3-105 was used in immunohistochemistry to study the role of NMDAR signaling in tumorigenesis using a murine model
|Li L,Hanahan D||Cell (153:86)||2013|