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- Peptide (C)TTKINMDDLQPSENEDKS, corresponding to amino acid residues 848-865 of rat CaV1.2 (Accession P22002). Intracellular loop between domains II and III.
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Western blot analysis of rat brain membrane:1. Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001), (1:200).
2. Guinea pig Anti-CaV1.2 (CACNA1C) Antibody, preincubated with the negative control antigen.
Western blot analysis of CaV1.2-transfected Xenopus oocytes (lane 1) and non-transfected oocytes lysates (lane 2):1. Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001), (1:200) in CaV1.2 (CACNA1C) Channel Overexpressed Membrane Fractions (LX-104).
2. Guinea pig Anti-CaV1.2 (CACNA1C) Antibody in non-transfected oocytes.
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Immuno-colocalization of CaV1.2 and GABA(A) α1 Receptor in rat cerebellumImmunohistochemical staining of rat cerebellum using Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001) and Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 Antibody (#AGA-001-AG). A. CaV1.2 (red) is detected mostly in Purkinje cells (arrow). B. In the same section, GABA(A) α1 Receptor (green) is observed in the granule layer. C. Merge of the two images suggests some colocalization between CaV1.2 and GABA(A) α1 Receptor in the rat granule layer but only CaV1.2 appears in Purkinje cells.
Expression of CaV1.2 in human atriaImmunohistochemical staining of human left atrium using Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001), (1:100).
The picture was kindly provided by Dr. Van Wagoner, D.R. from the Department of Molecular Cardiology, Cleveland Clinic, Cleveland, Ohio, USA. Lovano, B. and Peterson, J. collected the data.
Expression of CaV1.2 in rat heartImmunohistochemical staining of rat heart paraffin embedded sections using Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001). A. CaV1.2 staining (green) appears mainly in the cardiac muscle, and in a lesser intensity in the tunica intima layer of the smooth muscle of the muscular arteries. B. Nuclear staining using DAPI as the counter stain. C. Merged images of A and B.
Expression of CaV1.2 in mouse hippocampusImmunohistochemical staining of mouse dentate gyrus using Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001). A. CaV1.2 (green) appeared in the outer molecular layer of the dentate gyrus and in the granule layer. B. Counterstain with DAPI (blue) outlines the granule layer of the dentate gyrus.
- 1. Bauer, C.S. et al. (2010) Curr. Opin. Neurobiol. 20, 563.
- 2. Arikkath, J. et al. (2003) Curr. Opin. Neurobiol. 13, 298.
- 3. Catterall, W.A. (2000) Annu. Rev. Cell. Dev. Biol. 16, 521.
- 4. Davies, A. et al. (2007) Trends Pharmacol. Sci. 28, 220.
- 5. Dai, S. et al. (2009) Physiol. Rev. 89, 411.
- 6. Zuccotti, A. et al. (2011) Trends Pharmacol. Sci. 32, 366.
- 7. Hell, J.W. et al. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 3362.
- 8. De Jongh, K.S. et al. (1996) Biochemistry 35, 10392.
- 9. Gao, T. et al. (2001) J. Biol. Chem. 276, 21089.
- 10. Liao, P. et al. (2010) Pflugers. Arch. 460, 353.
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Voltage-gated Ca2+ channels (CaV), enable the passage of Ca2+ ions in a voltage dependent manner. These heteromeric entities are formed in part by the pore-forming α1 subunit which determines the biophysical and pharmacological properties of the channel1.
L-type Ca2+ channels make up one of three voltage-gated Ca2+ channel families. Four different α1 isoforms (CaV1.1 to CaV1.4) belong to the L-type subfamily. Structurally, each α1 subunit has four homologous domains (I-IV) and each domain has a six transmembrane section. Like many other voltage-gated channels, L-type Ca2+ channels have auxiliary subunits which are responsible for modulating the surface expression and properties of the channels2-5.
CaV1.1 is mostly expressed in the skeletal muscle, while CaV1.4 is mainly detected in the retina. The expression of both CaV1.2 and CaV1.3 is more extensive and includes neurons, heart, smooth muscle, inner ear, retina and pancreas6. L-type Ca2+ channels are involved in and modulate a variety of physiological functions such as muscle contraction, hormone secretion, neuronal excitability and gene expression5.
CaV1.2 undergoes various post-translational modifications. For example, it can undergo proteolytic cleavage at its C-terminal. This cleavage has been shown to take place in neurons following the activation of NMDA receptors5,7 and in the heart5,8,9. The cleaved moiety can still interact with the channel and its general purpose is to modulate channel activity5. Other postranslation modifications of the channel include phosphorylation of CaV1.2 by a number of kinases such as PKA, PKC, Src and CaMKII5. In addition, it is not surprising that phosphatases also regulate channel activity, as they are required to antagonize the activity of the various kinases known to phosphorylate CaV1.2 5.
The fact that CaV1.2 plays a prominent role in proper cardiac function has prompted endless studies regarding its regulation. Such studies have concluded that dysregulation of the channel leads to anomalies in heart contraction and thus heart failure5. Likewise, CaV1.2 defects have been detected in autism and bipolar disorder10.
Immuno-colocalization of CaV1.2 and GABA(A) α1 Receptor in rat and mouse hippocampusImmunohistochemical staining of mouse and rat hippocampal dentate gyrus using Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001) and Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 Antibody (#AGA-001-AG) in the same section. Both CaV1.2 (red) and GABA(A) α1 Receptor (green) are detected in neuron-shaped cells (arrows). Staining suggests partial colocalization between CaV1.2 and GABA(A) α1 Receptor in a sub-population of dentate gyrus neurons. A. Mouse hippocampus. B. Rat hippocampus.
Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001), raised in guinea pigs, is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot and immunohistochemistry applications. It and has been designed to recognize CaV1.2 from mouse, rat and human samples. The antigen used to immunize guinea pigs is the same as Anti-CaV1.2 (CACNA1C) Antibody (#ACC-003) raised in rabbit. Our line of guinea pig antibodies enables more flexibility with our products such as immuno-colocalization studies, immunoprecipitation, etc.
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【过敏性咳嗽血清学IGE检查】
以过敏患者血清作为实验材料的本外试验方法称为血清学试验。其它体液如炎症部位分泌物、渗出物、灌洗液也可采用相同的实验方法进行检测。主要检测项目有总IgE和特异性IgE，即过敏原特异性IgE。
【什么是总IgE？】
IgE即免疫球蛋白E，是I型变态反应病如过敏性鼻炎、过敏性哮喘、异位性皮炎、湿疹、急慢性荨麻疹发病机制中起主要作用的免疫分子，因而在过敏反应的免疫学实验诊断中是首选的检测项目。总IgE是过敏性疾病的特异性检查项目，IgE水平增高提示I型变态反应病的可能性大，但不能用于判断过敏原。
【IgE的特点】
IgE是血清浓度最低的免疫球蛋白 ，只有血清中IgG浓度的万分之一。IgE对热不稳定，是半衰期最短一的免疫球蛋白 ，只有2.8天，与细胞表面结合的IgE半衰期稍长，8~14天，IgE由变应原入侵部位（鼻咽、支气管、胃肠道）的黏膜固有层中的浆细胞合成。在各类免疫球蛋白中，IgE是合成率最低、分解率最高的。属于亲细胞抗体，过敏体质者的胎儿脐带血中IgE浓度可能升高，检测脐血中IgE浓度可用于评估胎儿过敏体质的可能性。
【IgE检测方法】
通常用ELISA方法检测总IgE。由于血清IgE浓度很低，一般酶免疫试验方法的敏感性不足以检出血清IgE，现在常规实验室检测血清IgE的试剂盒采用生物素——抗生物素蛋白 放大的ELISA。试剂盒中所含用于制定标准曲线的IgE标准品和检测结果的IgE浓度单位与其它免疫球蛋白 不同，不是用mg/L表示，而是用u/ml或ku/l表示。
【IgE的正常值（参考范围）】：
血清IgE水平在正常人群中呈偏态分布，即多数人为0或接近于0，IgE水平越高的人数越少。因此计算平均值时应计算几何平均值才能反映其真实情况，即用对数转换后其分布才能近似正态分布。
健康人群血清IgE水平与年龄关系较大，小儿和老年人的IgE水平低于成年人。新生儿血清中IgE水平很低，接近于零。随年龄增长，IgE水平也不断升高，5~7岁后接近正常人水平。按Pharmacia公司提供的参考范围，1个月以内<12KU/L,1岁<11KU/L,2~4岁<33KU/L,5岁以上至成人<85KU/L.
过敏性疾病患者的血清IgE水平可达2000~8000KU/L,当IgE水平高于2000KU/L时应考虑寄生虫感染.
有时血清总IgE水平检测结果为0或参考范围内低值,并不能排除过敏性疾病的可能,须结合临床表现和血清特异性IgE检测结果进行判断.
【什么是特异性IgE检测(sIgE）？】
通常所称的过敏原检测，并非真正检测血液样本中的过敏原分子，而是间接地检测其中针对某种过敏原的特异性IgE分子，特异性IgE检测实际上是检测过敏原特异性IgE，即检测样本中针对某种变应原的特异性IgE，从而间接地判断患者是否对某种过敏原过敏。
环境中常见的过敏原包括以下类别：
寄生虫和微生物：各种螨类（屋尘螨和粉尘螨等）、各种真菌（点青霉、烟曲霉、分枝孢霉、交连孢霉等）、蟑螂。
植物花粉：各种草花粉（豚草、葎草、蒿草）、各种树花粉（桑树、柏树、悬铃木、桦树、榆树、柳树、杨树等）。
动物皮毛：猫、狗、马、鸽子等动物的毛和皮屑。
也就是说只要是兔子产生的抗体，羊抗兔（也叫抗抗体或者2抗）都可与之结合。
兔子产生的是一级抗体
羊的为二级抗体
在试验中，二抗经常被做了手脚，打上了标记，比如辣根过氧化物酶，经常被用在ELISA，WESTERN等经典的生物学试验中。
