FLIP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:FLIP抗体
中文名称凋亡调节基因之一抗体
别 名CASP8 and FADD-like apoptosis regulator subunit p43; CASP8 and FADD-like apoptosis regulator subunit p43; Flice-like Inhibitory protein; c FLIP; c FLIPL; c FLIPR; c FLIPS; c-FLIP; CASH; CASP8 and FADD like apoptosis regulator; CASP8 and FADD like apoptosis regulator precursor; CASP8AP1; Caspase Eight Related Protein; Caspase homolog; Caspase Homologue; Caspase Like Apoptosis Regulatory Protein; Caspase related inducer of apoptosis; CASPER; Cellular FLICE like inhibitory protein; CFLA; CFLAR; CLARP; FADD like anti apoptotic molecule; FADD Like Anti-apoptotic Molecule 1; FADD-like antiapoptotic molecule 1; FADD like antiapoptotic molecule 1; FADD Like Apoptosis Regulator; FLAME 1; FLAME; FLAME1; FLAME-1; FLICE Inhibitor Protein; FLIP; FLIPs; I FLICE; I-FLICE; Inhibitor of FLICE; Inhibitor of FLICE; MACH Related Inducer of Toxicity; MACH-related inducer of toxicity; mFLIP; MRIT; USURPIN; Usurpin beta; FLICE-like inhibitory protein short form; FLICE-like inhibitory protein long form; CFLAR_HUMAN; Cellular FLICE-like inhibitory protein.
说 明 书50ul 100ul 200ul
研究领域肿瘤 细胞生物 信号转导 细胞凋亡
抗体来源Rabbit
克隆类型Polyclonal
交叉反应 Human, Mouse, Rat, Dog, Pig, Cow, Rabbit,
产品应用WB=1:500-2000 ELISA=1:500-1000 IHC-P=1:400-800 IHC-F=1:400-800 IF=1:100-500 (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.
FLIP抗体分 子 量43/52kDa
细胞定位细胞浆
性 状Lyophilized or Liquid
浓 度1mg/1ml
免 疫 原KLH conjugated synthetic peptide derived from human CASP8 and FADD-like apoptosis regulator subunit p43
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.
FLIP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。
FLIP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!