beta-Actin (Loading Control)抗体做wb时,一抗和二抗浓度是怎么确定的?怎么设置抗体滴度:
一抗的用量和样品中的目标分子含量相关,如果含量较高,就不需要用太多一抗,以免浪费和条带太亮不易分析。推荐采用厂家的抗体浓度(义翘神州会提供抗体用量、天然细胞株检测结果),如果样品与厂家的不是一种细胞,可以尝试上下摸索一两个浓度梯度;
二抗的用量,和你所在实验室常规的用量就可以,如果出现效果不理想的状态,二抗的作用可以通过阳性对照和阴性对照、以及其他人的使用效果看出来有没有问题。
英文名称:beta-Actin (Loading Control)抗体
中文名称β-肌动蛋白/β-Actin 抗体(内参抗体)
别 名Beta Actin; beta-Actin; ACTB; Actin cytoplasmic 1; Actin, beta; Beta actin; beta cytoskeletal actin;A X actin like protein; ACTB; Actin cytoplasmic 1; alpha sarcomeric Actin; Actx; Beta cytoskeletal actin; Melanoma X actin; PS1TP5BP1; ACTB_HUMAN.
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说 明 书50ul 100ul 200ul
产品类型内参抗体
研究领域肿瘤 细胞生物 信号转导 细胞骨架
抗体来源Rabbit
克隆类型Polyclonal
交叉反应 Human, Mouse, Rat, Chicken, Dog, Pig, Cow, Rabbit, Sheep, Bee, Fish, Guinea Pig, Hamster, Goat, Cat, mt,op
产品应用WB=1:5000-20000 ELISA=1:5000-20000 IHC-P=1:500-1000 IHC-F=1:500-1000 Flow-Cyt=1μg/Test IF=1:100-1000 (石蜡切片需做抗原修复)
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
beta-Actin (Loading Control)抗体分 子 量42kDa
细胞定位细胞浆
性 状Lyophilized or Liquid
浓 度1mg/1ml
免 疫 原Synthetic MAP peptide derived from human beta-Actin
亚 型IgG
纯化方法affinity purified by Protein A
储 存 液0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
保存条件Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C
beta-Actin (Loading Control)抗体ELISA如何选一抗与二抗:
一抗是与底物相结合的,同时一抗必须有抗原识别位点,能被另一种抗体识别并结合。
二抗是能与一抗的抗原识别位点识别并结合,同时,二抗必须有可以观测、计量的性质。
这些性质可以是:
①二抗本身带有颜色,可以通过分光度仪检测到二抗的浓度;
②二抗还可以作为一种酶,催化另一种底物,通过检测底物或者生成物的浓度检测出二抗的浓度;等等……有很多,就是说必须是可测量的。
beta-Actin (Loading Control)抗体保存得当与否,直接决定了抗体的活性和使用效果!如果抗体保存得当,大部分抗体活性都可以维持数月甚至数年。收到抗体后请务必在12000rpm离心20-30s后再打开管盖进行分装和保存!抗体使用非常特殊的储存管,只有在12000rpm离心20-30s才能将附着在管壁或盖内的抗体全部离心下来。即使是在10000rpm离心也会导致离心不完全,导致出现抗体的量不足的现象!