WB
Western blot analysis using TFEB antibody against Hepg2(1), A549(2), A431(3), Hek293(4), Mcf-7(5) cell. 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 1:2000, 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.WB
Western blot analysis using TFEB antibody against Mouse skin tissue(1), Rat stomach tissue(2). 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 1:2000, 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 breast cancer tissues using TFEB 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 esophageal cancer tissues using TFEB 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 TFEB 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 renal carcinoma tissues using TFEB 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 HUVEC cells using TFEB 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 15 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.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 |
| 来源 | 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 | TFEB |
| 基因别名 | ALPHATFEB; BHLHE35; TCFEB |
| 基因全称 | transcription factor EB |
| 基因简介 | Enables DNA-binding transcription factor activity; enzyme binding activity; and transcription cis-regulatory region binding activity. Involved in several processes, including antibacterial innate immune response; cellular response to amino acid starvation; and positive regulation of metabolic process. Located in cytosol and lysosomal membrane. Is active in nucleus. [provided by Alliance of Genome Resources, Jul 2025] |
| 蛋白别名 | Alpha TCFEB; basic helix-loop-helix leucine zipper (bHLH-ZIP) transcription factor; class E basic helix-loop-helix protein 35; RP4-696P19.3; T-cell transcription factor EB; TFEB protein |
| 分子量 | 53kDa |
| Uniprot ID | P19484 |
| 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 | Nucleus, Cytoplasm, cytosol, Lysosome membrane |
| 产品形态 | Liquid |
| 缓冲液 | Purified antibody in PBS with 0.05% PC300. |
| 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。
