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
Immunocytochemical labeling of phosphorylated actin in paraformaldehyde fixed and acetone permeabilized C2C12 cells treated with pervanadate. The cells were labeled with mouse monoclonal anti-Actin (AM2021) and rabbit polyclonal anti-Actin (AP1651) or anti-Actin (Tyr-53) (AP1671). The antibodies were detected using Goat anti-Mouse DyLight® 594 or Goat anti-Rabbit DyLight® 488.
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 splenocytesAP1671 Vonach, C. et al. (2011) British J Cancer. 105:263. WB: human endothelial cellsThis kit contains:
| CATALOG# | DESCRIPTION | SIZE | APPLICATIONS | SPECIES REACTIVITY | ICC DILUTION |
AP1651 | Actin (N-terminal region) Rabbit pAb | 50 μl | WB, E, IP, ICC, IHC | Hu, Rt, Ms, Ck | 1:50 |
AP1671 | Actin (Tyr-53), phospho-specific Rabbit pAb | 50 μl | WB, E, ICC | Hu, Rt, Ms, Ck | 1:50 |
AM2021 | Actin (C-terminal region) Mouse mAb | 50 μl | WB, E, ICC, IHC | Hu, Rt, Ms, Ck | 1:50 |
MS3031 | Anti-Mouse Ig:DyLight®594 Goat pAb | 100 μl | ICC, IHC | Ms | 1:200 |
RS3261 | Anti-Rabbit Ig:DyLight®488 Goat pAb | 100 μl | ICC, IHC | Rb | 1:200 |
KIT SUMMARY
The actin phospho-regualtion kit can be used to label actin phosphorylation at Tyr-53 relative to total actin expression pattern. The kit contains anti-Actin (Tyr-53) phospho-specific antibody, and two antibodies for detecting total actin. In addition, secondary reagents conjugated to DyLight® 488 or DyLight® 594 are included for labeling primary antibodies for detection using green (493Ex/518Em) or red (593Ex/618Em) filter sets.
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请各位高手指点一下小弟吧！
要用几种不同材料来做这个实验
还要自己设计试验方法步骤所用仪器用品！
实在很是头疼谢谢！
许多食品属于乳胶体（冰淇淋、豆奶），蛋白质成分在稳定这些胶态体系中通常起着重要的作用。天然乳胶体靠脂肪球“这种“膜”由三酰甘油、磷脂、不溶性脂蛋白和可溶性蛋白的连续吸附层所构成。蛋白质一般对水/油（W/O）型乳胶液的稳定性较差。这可能是因为大多数蛋白质的强亲水性使大量被吸附的蛋白质分子位于界面的水相一侧。蛋白质的表面活性不仅与蛋白质中氨基酸的组成、结构、立体构象、分子中极性和非极性残基的分布与比例，二硫键的数目与交联，以及分子的大小、形状和柔顺性等内在因素有关，而且与外界因素，甚至加工操作有关。凡是能影响蛋白质构象和亲水性与疏水性的环境因素，诸如pH、温度、离子强度和盐的种类、界面的组成、蛋白质浓度、糖类和低分子量表面活性剂，能量的输入，甚至形成界面加工的容器和操作顺序等，都将影响蛋白质的表面活性。
2．起泡性
食品泡沫通常是气泡在连续的液相或含可溶性表面活性剂的半固相中形成的分散体系。种类繁多的泡沫其质地大小不同，例如蛋白质酥皮、蛋糕、棉花糖和某些其他糖果产品、点心顶端配料、冰淇淋、蛋奶酥、啤酒泡沫、奶油冻和面包等。大多数情况下，气体是空气或CO2，连续相是含蛋白质的水溶液或悬浊液。某些食品泡沫是很复杂的胶态体系，例如冰淇淋中存在分散的和群集的脂肪球（多数是固体）、乳胶体（或悬浊液）、分散的冰晶悬浮体，多糖凝胶、糖和蛋白质的浓缩溶液以及空气气泡。各种泡沫的气泡大小不相同，直径从1微米到几cm不等，气泡的大小取决于多种因素，例如，液相的表面张力和粘度、输入的能量，分布均匀的细微气泡可以使食品产生稠性、细腻和松软性，提高分散性和风味感。
3.凝胶性
变性的蛋白质分子聚集并形成有序的蛋白质网络结构过程称为胶凝作用。胶凝是蛋白 质的重要功能性质，在许多食品的制备中起着主要作用，包括各种乳品、果冻、凝结蛋白、明胶凝胶、各种加热的碎肉或鱼制品、大豆蛋白质凝胶、膨化或喷丝的组织化植物蛋白和面包面团的制作等，中国人喜爱的豆腐食品，就是大豆蛋白胶凝作用的产物。蛋白质胶凝作用不仅可用来形成固态粘弹性凝胶，而且还能增稠，提高吸水性和颗粒粘结、乳状液或泡沫的稳定性。
4．溶解度
大豆蛋白质在溶解状态下才能发挥其在食品体系中的功能特性。大豆蛋白质的溶解度是指大豆蛋白质以胶体的形式分散到水中的能力。蛋白质分子的极性表面和所带的净电荷有助于分散体系的稳定。大豆蛋白质的溶解度可以用可溶性氮指数(NSI)和蛋白质分散度指数(PDI)两种方法表示。影响大豆蛋白质溶解度的因素主要包括温度、pH和无机盐。
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！
转谷氨酰胺酶(TG)、过氧化酶(POD)、多酚氧化酶(PPO)交联
戊二醛，便宜，但有毒性
EDC，无毒性，但较贵
SMCC ,DSS,DST MBS,SPDP
