Actin is a major cytoskeletal protein involved in diverse cellular functions including cell motility, adhesion, and morphology. Six different actin isoforms have been identified in vertebrates. There are four α isoforms: skeletal, cardiac, and two smooth muscle (enteric and aortic) actins, along with two cytoplasmic actins (β and γ). Actin exists in two principal forms, globular, monomeric (G) actin, and filamentous polymeric (F) actin. The assembly and disassembly of actin filaments, and also their organization into functional networks, is regulated by a variety of actin-binding proteins (ABPs). Phosphorylation may also be important for regulating actin assembly and interaction with ABPs. In Dictyostelium, phosphorylation of Tyr-53 occurs in response to cell stress and this phosphorylation may alter actin polymerization. In B cells, SHP-1 tyrosine dephosphorylation of actin leads to actin filament depolymerization following BCR stimulation.
References
Western blot analysis of mouse C2C12 cells untreated (lanes 1 & 3), or treated with pervanadate (1 mM) for 30 min (lanes 2 & 4). The blot was probed with anti-Actin (N-terminal) antibody (lanes 1 & 2) or anti-Actin (Tyr-53) antibody (lanes 3 & 4).
Formalin fixed, citric acid treated parafin sections of E18 mouse skeletal muscle. Sections were probed with anti-Actin (AM2021) then anti-Mouse:HRP before detection using DAB. (Images provided by Carl Hobbs and Dr. Pat Doherty at King"s College London).
The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.
*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blotmobilities of known proteins with similar MW.
Product References:
AM2021 Dutta, P et al. (2014) J Neurochem. 130(3):360 WB: rat brainAM2021 Muirhead, G et al. (2014) J Mol Neurosci. 53(1):125 WB: rat brainAM2021 Pritchard, AJ et al. (2014) PLoS One. 9(6):e99444 WB: mouse splenocytesRS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblastsAP1671 Vonach, C. et al. (2011) British J Cancer. 105:263. WB: human endothelial cellsMS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western BlotRS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western BlotThis kit contains:
| CATALOG# | DESCRIPTION | SIZE | APPLICATIONS | SPECIES REACTIVITY | MW (kDa) |
AP1651 | Actin (N-terminal region) Rabbit pAb | 50 μl | WB, E, IP, ICC, IHC | Hu, Rt, Ms, Ck | 42 |
AP1671 | Actin (Tyr-53), phospho-specific Rabbit pAb | 50 μl | WB, E, ICC | Hu, Rt, Ms, Ck | 42 |
AM2021 | Actin (C-terminal region) Mouse mAb | 50 μl | WB, E, ICC, IHC | Hu, Rt, Ms, Ck | 42 |
MS3001 | Anti-Mouse Ig:HRP Donkey pAb | 100 μl | WB, E | Ms | |
RS3251 | Anti-Rabbit Ig Light-Chain Specific:HRP Mouse mAb | 100 μl | WB, E, ICC, IHC | Rb |
KIT SUMMARY
The actin phospho-regulation kit can be used to examine phosphorylation of Ty-53 compared to the total expression of actin. The kit includes rabbit polyconal and mouse monoclonal antibodies to detect total actin, and secondary reagents for rabbit polyclonal and mouse monoclonal antibody detection.
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请各位高手指点一下小弟吧！
我最近在做磁珠与蛋白的偶联，想利用测上清液中剩余的蛋白量来检测偶联率，但是我发现交联剂对BCA法检测上清蛋白含量有显然影响，不知道有没有师兄师姐做过，最后是怎么解决偶联率检测的问题的。求教，谢谢！
甲苯二异氰酸酯（TDI）有两种异构体：2，4－甲苯二异氰酸酯和2，6－甲苯二异氰酸酯。甲苯二异氰酸酯是水白色或淡黄色液体，具有强烈的刺激性气味，在人体中具有积聚性和潜伏性，对皮肤、眼睛和呼吸道有强烈刺激作用，吸入高浓度的甲苯二异氰酸酯蒸气会引起支气管炎、支气管肺炎和肺水肿；液体与皮肤接触可引起皮炎。液体与眼睛接触可引起严重刺激作用，如果不加以治疗，可能导致永久性损伤。长期接触甲苯二异氰酸酯可引起慢性支气管炎。对甲苯二异氰酸酯过敏者，可能引起气喘、伴气喘、呼吸困难和咳嗽。
与乙醚、二甘醇、丙酮、四氯化碳、苯、氯苯、煤油、橄榄油混溶。能与含羟基的化合物、水、胺和具有活泼氢原子的化合物反应生成氨基甲酸酯、脲、氨基脲等。甲苯用混酸硝化得到2,4-和2,6-二硝基甲苯，然后在镍催化剂存在下加氢还原得到2,4-和2,6-二氨基甲苯，再在氯苯溶液中与光气反应制得。主要作为聚氨酯树脂的生产原料，用于生产聚氯酯泡抹塑料、涂料、橡胶、粘合剂、密封剂等。也可用作橡胶硫化剂、蛋白质交联剂等。包括泡沫塑料；聚氨酯涂料；聚氨酯橡胶；聚酰亚胺纤维和胶粘剂等也有一些应用。
明胶与干酪素都是蛋白质类，其分子结构同属氨基酸，但前者比后者价格便宜很多，而且在我国的产盆很大，因此具有代替干酪素做涂料胶粘剂的可能性。
1.用这个人类膜蛋白c端末短肽（14个残基，478-492）偶联KLH生产的兔多克隆抗体（abcam公司产品）
2.用大鼠的同源膜蛋白c端（具体哪一段不清楚）生产的兔多克隆抗体（abcam公司产品），我比对过人和大鼠的这个膜蛋白，同源性很高，c端完全一样（这里也很困惑公司为什么说是大鼠来源的）
3.用这个人类膜蛋白两个跨膜区之间的一个loop环序列（42个残基，218-260）生产的兔多克隆抗体（santa公司产品）
这三个抗体理论上来讲哪个更好些呢？请有经验的大虾给予指点！
另：我听说abcam的抗体比santa好，是吗？
在线等，急！
我现在做关于细胞膜分子交联，然后用western检测多聚体的实验
遇到很大的困难
刚开始用真核细胞做，就发现细胞交联后提取蛋白，比未交联的细胞蛋白总量减少
后来用原核先提取蛋白做实验
发现交联终止后，做western交联的孔曝光没有条带，后来用同样的方法重新做了一次PAGE，染胶，发现蛋白不见了
为什么交联后蛋白减少或是不见了呢
我用的都是Pierce公司的交联剂，用过EGS和BOSCOES，都有同样的问题
方法是综合说明书和一片文献上来的
到底问题出在哪里呢
请各位大侠帮帮忙
实验卡在这里进行不下去了
我快急死了
请有经验的，用过交联剂的老师，同学多多指导
万分感谢！！
