WB
Western blot analysis of lysates from HeLa cells using 4-Hydroxynonenal Rabbit mAb at 1:10000 dilution incubated overnight at 4℃. HeLa cells were treated with 4-HNE (0.2 mg/ml) at 37℃ for 30 minutes. Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L)at 1:10000 dilution. Lysates/proteins: 30 μg per lane. Blocking buffer: 3 % nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 30s.ICC/IF
Confocal imaging of HeLa cells (treated with 4-HNE) and HeLa cells (untreated) using 4 Hydroxynonenal Rabbit mAb (dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (dilution 1:400) followed by incubation with TSZ® 488 Goat Anti-Mouse IgG(H&L) Ab (dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.IF-P
Confocal imaging of paraffin-embedded Mouse brain and 5xFAD Mouse brain tissue using 4-Hydroxynonenal Rabbit mAb (dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IF staining. Objective: 40x.FC
Flow cytometry: 1X10^6 HeLa cells (negative control,left) and HeLa cells (treated with 4-Hydroxynonenal,right) were intracellularly-stained with 4-Hydroxynonenal Rabbit mAb (2 μg/mL,orange line) or Rabbit IgG isotype control (2 μg/mL,blue line), followed by FITC conjugated goat anti-Rabbit pAb staining. Non-fluorescently stained cells were used as blank control (red line).| 抗体类型 | Primary Antibodies |
|---|---|
| 免疫原 | Chemical compounds corresponding to 4-Hydroxynonenal. |
| 克隆性 | monoclonal |
| 同种型 | IgG |
| 宿主 | Rabbit |
| 已测试的应用 | FCICC/IFIF-PWB |
| 物种反应性 | All |
| 浓度 | 1mg/ml |
| 纯化 | Affinity purified |
| 蛋白别名 | 4-HNE |
| 分子量 | 42-350kDa |
| 应用稀释比 |
WB:1:1000-1:10000 ICC/IF:1:200-1:500 IF-P:1:200-1:500 FC:1:100-1:500 |
| 亚细胞定位 | Cell membrane, Cytoplasm, Nucleus. |
| 产品详情 | 4-hydroxy-2-nonenal (4-hydroxynonenal, 4-HNE) is a highly reactive aldehyde generated by the exposure of polyunsaturated fatty acids to peroxides and reactive oxygen species (ROS). It non-enzymatically forms stable protein adducts with histidine, lysine, and cysteine side chains that have been used as biomarkers for oxidative damage in cells. Conditions where 4-HNE immunoreactivity has been observed include include inflammation, neurodegenerative diseases, and ischemic damage to the heart and brain. |
| 产品形态 | Liquid |
| 缓冲液 | Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3. |
| 保存条件 | Store at -20℃. Avoid freeze / thaw cycles. |
请在以下方框中输入您的动物实验信息后点击计算，可以得到母液配置方法和体内配方的制备方法：
比如您的给药剂量是10 mg/kg，每只动物体重20 g，给药体积100 μL， 一共给药动物10只，您使用的配方为 10% DMSO + 40% PEG300 + 5% Tween 80 + 45% Saline / PBS / ddH2O， 那么您的工作液浓度为2 mg/mL。
母液配置方法：2 mg 药物溶于 100 μL DMSO （ 母液浓度为 20 mg/mL ）, 如您需要配置的浓度超过该产品的溶解度，请先与我们联系。
体内配方的制备方法： 取 100 μL DMSO 母液， 添加 400 μL PEG300 混匀澄清， 再加 50 μL Tween 80, 混匀澄清, 再加 450 μL Saline / PBS / ddH2O 混匀澄清
以上为“体内实验配液计算器”的使用方法举例，并不是具体某个化合物的推荐配制方式，请根据您的实验动物和给药方式选择适当的溶解方案。
方案所需的各类助溶剂如： DMSO， PEG300， PEG400， Tween 80， SBE-β-CD， 玉米油等， 均可在诺渊网站点击购买。
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL Saline/PBS/ddH2O，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
