WB
Western blot analysis using AMPK alpha antibody against K562(1), HepG2(2) cell lysate. 12% SDS-PAGE gel. Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane, and block it with TBST plus skimmed milk powder for one hour. Dilute the primary antibody with the antibody diluent at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST, 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP, at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST again, 5 minutes each time. Use ECL for luminescence. staining time: 60S.IHC
Immunohistochemical analysis of paraffin-embedded Human lung cancer tissues using AMPK alpha antibody with DAB staining. Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6. 0) for 2 minutes. Wash the sections with ddH₂O and PBS. Block the tissue with 10% non-immune goat serum at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP, at a ratio of 1:200 and incubate for one hour. Use DAB as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.ICC/IF
Immunofluorescence analysis of A549 cells using AMPK alpha antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with TSZ® 594-Phalloidin. Cells are fixed in 4% paraformaldehyde at room temperature for 20 minutes. Then, they are permeabilized with a PBS solution containing 0. 1% Triton X-100 at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum at room temperature for 1 hour. The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, TSZ® 488 Goat Ant-Rabbit IgG(H&L) (Green) is diluted at a ratio of 1:400 and incubated with the cells for 1 hour. Nuclear DNA is labeled with DAPI (Blue). F-actin is stained with TSZ® 594-Phalloidin (Red), diluted 1:100 for 30 minutes.IHC
Immunohistochemical analysis of paraffin-embedded Human kidney tissues using AMPK alpha antibody with DAB staining. Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6. 0) for 2 minutes. Wash the sections with ddH₂O and PBS. Block the tissue with 10% non-immune goat serum at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP, at a ratio of 1:200 and incubate for one hour. Use DAB as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissues using AMPK alpha antibody with DAB staining. Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6. 0) for 2 minutes. Wash the sections with ddH₂O and PBS. Block the tissue with 10% non-immune goat serum at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP, at a ratio of 1:200 and incubate for one hour. Use DAB as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded Human rectal cancer tissues using AMPK alpha antibody with DAB staining. Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6. 0) for 2 minutes. Wash the sections with ddH₂O and PBS. Block the tissue with 10% non-immune goat serum at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP, at a ratio of 1:200 and incubate for one hour. Use DAB as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.ELISA
Coat the plate with immunizing antigen. Blue line：Immune serum; Orange line: Pre-immune serum.| 抗体类型 | Primary Antibodies |
|---|---|
| 免疫原 | Polypeptide |
| 克隆性 | Polyclonal |
| 同种型 | IgG |
| 宿主 | Rabbit |
| 已测试的应用 | ELISAICC/IFIHCWB |
| 物种反应性 | HumanMouseRat |
| 浓度 | 1mg/ml |
| 纯化 | Protein A |
| 基因名称 | PRKAA1 |
| 基因别名 | AMPK; AMPKa1; AMPK alpha 1 |
| 基因全称 | protein kinase AMP-activated catalytic subunit alpha 1 |
| 基因简介 | The protein encoded by this gene belongs to the ser/thr protein kinase family. It is the catalytic subunit of the 5'-prime-AMP-activated protein kinase (AMPK). AMPK is a cellular energy sensor conserved in all eukaryotic cells. The kinase activity of AMPK is activated by the stimuli that increase the cellular AMP/ATP ratio. AMPK regulates the activities of a number of key metabolic enzymes through phosphorylation. It protects cells from stresses that cause ATP depletion by switching off ATP-consuming biosynthetic pathways. Alternatively spliced transcript variants encoding distinct isoforms have been observed. [provided by RefSeq, Jul 2008] |
| 蛋白别名 | 5 AMP activated protein kinase alpha 1catalytic subunit antibody;5 AMP activated protein kinase catalytic alpha 1 chain antibody;5' AMP activated protein kinase catalytic subunit alpha 1 antibody;5'-AMP-activated protein kinase catalytic subunit alpha-1 antibody;AAPK1 antibody;AAPK1_HUMAN antibody;ACACA kinase antibody;acetyl CoA carboxylase kinase antibody;AI194361 antibody;AI450832 antibody |
| 分子量 | 62 kDa, 64 kDa |
| Uniprot ID | Q13131;1P54646 |
| 应用稀释比 |
WB:1:1000-1:2000 IHC:1:200-1:2000 ICC/IF:1:100-1:500 |
| 修饰 | Unmodification |
| 标记 | Unconjugated |
| 亚细胞定位 | Cytoplasm; Nucleus |
| 产品形态 | Liquid |
| 缓冲液 | Purified antibody in PBS with 0.05% sodium azide. |
| 保存条件 | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
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。
