This product is freeze dried. All water molecules have been removed.
This antibody is shipped with its antigen FREE of charge!
- Peptide GHSHDVTERELRN(C), corresponding to amino acid residues 41-53 of rat NR2A (Accession Q00959). Extracellular, N-terminus.
- Expression of NR2A in mouse brain sectionsImmunohistochemical staining of perfusion-fixed frozen mouse brain sections using Anti-NMDAR2A (GluN2A) (extracellular)-ATTO-488 Antibody (#AGC-002-AG), (1:80). A. NR2A staining (green) in striatum is detected in neuronal profiles. B. NR2A staining in cingulate cortex is also detected in neuronal profiles. Cell nuclei are labeled with DAPI (blue).
- 1. Dingledine, R. et al. (1999) Pharmacol. Rev. 51, 7.
- 2. Mayer, M.L. and Armstrong, N. (2004) Annu. Rev. Physiol. 66, 161.
- 3. Prybylowski, K. and Wenthold, R.J. (2004) J. Biol. Chem. 279, 9673.
- 4. Mayer, M.L. (2006) Nature 440, 456.
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The NMDA receptors are members of the glutamate receptor family of ion channels that also include the AMPA and Kainate receptors.
The NMDA receptors are encoded by seven genes: one NMDAR1 (or NR1) subunit, four NR2 (NR2A-NR2D) and two NR3 (NR3A-NR3B) subunits. The functional NMDA receptor appears to be a heterotetramer composed of two NMDAR1 and two NMDAR2 subunits. Whereas the NMDAR2 subunits that assemble with the NMDAR1 subunit can be either of the same kind (i.e. two NMDAR2A subunits) or different (one NMDAR2A with one NMDAR2B). NMDAR3 subunits can substitute the NMDAR2 subunits in their complex with the NMDAR1 subunit.
NMDAR is unique among ligand-gated ion channels in that it requires the simultaneous binding of two obligatory agonists: glycine and glutamate that bind to the NMDAR1 and NMDAR2 binding sites respectively. Another unique characteristic of the NMDA receptors is their dependence on membrane potential. At resting membrane potentials the channels are blocked by extracellular Mg2+. Neuronal depolarization relieves the Mg2+ blockage and allows ion influx into the cells. NMDA receptors are strongly selective for Ca2+ influx differing from the other glutamate receptor ion channels that are non-selective cation channels.
Ca2+ entry through the NMDAR regulates numerous downstream signaling pathways including long term potentiation (a molecular model of memory) and synaptic plasticity that may underlie learning. In addition, the NMDA receptors have been implicated in a variety of neurological disorders including epilepsy, ischemic brain damage, Parkinson’s and Alzheimer’s disease.
NMDA receptors expression and function are modulated by a variety of factors including receptor trafficking to the synapses and internalization as well as phosphorylation and interaction with other intracellular proteins.
Immuno-colocalization of GluN2A and SynGAP in rat cingulate cortex.Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-NMDAR2A (GluN2A) (extracellular)-ATTO-488 Antibody (#AGC-002-AG), (1:200) and Anti-SynGAP Antibody (#APZ-032), (1:200), followed by donkey-anti-rabbit-Cy3. A. GluN2A staining (green) appears in neuronal profiles (arrows). B. SynGAP staining (red) is detected mostly in apical dendrites (horizontal arrows) and in neuronal soma (vertical arrows). C. Merge of the two images demonstrates colocalization in some neurons (vertical arrows). Cell nuclei are stained with DAPI (blue).
Anti-NMDAR2A (GluN2A) (extracellular) Antibody (#AGC-002) is a highly specific antibody directed against an extracellular epitope of the rat protein. The antibody can be used in western blot, immunoprecipitation, immunocytochemistry, and immunohistochemistry. It has been designed to recognize GluN2A from rat, mouse, and human samples.
Anti-NMDAR2A (GluN2A) (extracellular)-ATTO-488 Antibody (#AGC-002-AG) is directly labeled with an ATTO-488 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-488 label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC. Anti-NMDAR2A (GluN2A) (extracellular)-ATTO-488 Antibody has been tested in immunohistochemistry and is especially suited for experiments requiring simultaneous labeling of different markers.
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英 ['aɪsətaɪp]美 ['aɪsəˌtaɪp]
n. 同型动物(或植物),图形文字,象征性图像;同位型;同模;同号模式
同型对照(阴性对照)管:A同型+B同型+C同型
单阳性对照
A检测管:A+B同型+C同型
B检测管:B+A同型+C同型
C检测管:C+A同型+B同型
不存在荧光重叠的两个抗体不需要调节补偿。
样本为PBMC,想用三色标记Treg细胞,分别为CD4-PECY,CD25-Fitc,Foxp3-PE,想知道如何设置荧光补偿,用的是BD的流式仪,1、是设置阴性管一个,单阳性管每种一个,样品管一个么?
荧光素强度依次为PE, PECY5 APC FITC, PE最强,FITC最弱。 所以,建议抗体组合为:
Foxp3-PE,CD4-FITC,CD25-PECy5.5或者CD25-APC。
同时购买三个相应的同型对照抗体。
样品管设置如下:
1. cell + FITC-isotype + Pecy5.5( or APC)-isotype + PE-isotype
2. Cell + FITC-isotype + CD25-mAb + PE-isotype
3. Cell + PE-mAb + FITC-isotype + Pecy5.5( or APC)-isotype
4. Cell + CD4-FITC + Foxp3-PE + CD25 -PECy5.5( or APC)。 [这一管的数据可以拿来分析用]。
在充分了解调整电压和补偿原理的基础上,也可以做一些简化。如果不太懂,就按照这个来。其中Foxp3的抗体和同型对照抗体都是在固定破膜以后再加,不是与CD4或者CD25同时加。
比如你用anti-MHC-II,mouse IgG1-FITC,那同型就用非特异性的mouse IgG1-FITC,同样浓度
手头上,有eBioscience家的同型抗体,为PE标记(mouseIgG1,k),说明书写使用为2.5ug/test。
按照说明书的浓度加入后结果发现,同型对照抗体组比实验组和阴性对照组(不表达CD80)荧光强度要高,很明确我的同型对照加入的剂量多了,但正确的加法是多少?按照质量算吗?请高手指教,非常感谢!!
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