StemTAG™ Alkaline Phosphatase Activity Assay Kit, Colorimetric
(16 references)-
- 适用
- 人
- 检测方法
- Fluorometric, Colorimetric
- 应用范围
- Activity Assay (AcA)
- 品牌
- StemTAG™
- 样品类型
- Cell Samples
- 组件
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- StemTAG™ AP Activity Assay Substrate : One bottle - 5 mL
- Cell Lysis Buffer : One bottle - 20 mL
- 10X Stop Solution : One bottle - 10 mL
- AP Activity Assay Standard : One tube - 1 mL of 5 mM p-Nitrophenol
- 试剂未包括
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- Human or Mouse Embryonic Stem Cells and Culture Medium
- 1X PBS
- 1X PBST (1X PBS containing 0.05 % Tween-20)
- Deionized Water
- Light Microscope
- 96-well Microplate Reader
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Tumor Necrosis Factor alpha (TNF alpha) ELISA Kit
TNF alpha 适用: 大鼠 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Interleukin 1, beta (IL1B) ELISA KitIL1B 适用: 大鼠 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Tumor Necrosis Factor alpha (TNF alpha) ELISA KitTNF alpha 适用: 小鼠 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
High Mobility Group Box 1 (HMGB1) ELISA KitHMGB1 适用: 人 Colorimetric Sandwich ELISA 62.5 pg/mL - 4000 pg/mL Plasma, Serum
Lipopolysaccharides (LPS) ELISA Kit适用: Various Species Colorimetric Competition ELISA 12.35 ng/mL - 1000 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Tumor Necrosis Factor alpha (TNF alpha) ELISA KitTNF alpha 适用: 人 AA 77-233 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Plasma (EDTA), Plasma (citrate), Plasma (heparin), Serum
Interleukin 6 (IL6) ELISA KitIL6 适用: 人 Colorimetric Sandwich ELISA 7.8-500 pg/mL Cell Culture Supernatant, Plasma, Serum, Tissue Homogenate, Urine
Vascular Cell Adhesion Molecule 1 (VCAM1) ELISA KitVCAM1 适用: 人 Colorimetric Sandwich ELISA 1.563-100 ng/mL Plasma, Serum, Tissue Homogenate
Matrix Metalloproteinase 2 (MMP2) ELISA KitMMP2 适用: 人 Colorimetric Sandwich ELISA 0.78 ng/mL - 50 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Interleukin 1, beta (IL1B) ELISA KitIL1B 适用: 人 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
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- 应用备注
- Optimal working dilution should be determined by the investigator.
- 说明
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- Measure the ubiquitous alkaline phosphatase marker in embryonic stem cells and embryonic germ cells
- Quantify activity by colorimetric or fluorometric plate reader
- 试剂准备
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- 1X Stop Solution: Prepare a 1X Stop Solution by diluting the provided 10X stock 1:10 in deionized water. Store the diluted solution at room temperature.
- 实验流程
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- Culture mouse ES cells in medium containing LIF, alternatively, culture human ES cells on a MEF feeder layer.
- Gently aspirate the medium from the ES cells and wash the cells twice with cold PBS. Aspirate the wash solutions.
- Lyse the cells in Cell Lysis Buffer (0.5 mL for a 35 mm dish).
- Incubate for 10 minutes at 4 °C, remove the solution and spin down the cell debris at 12,000 X g for 10 minutes. Save the supernatant as cell lysate. Perform a BCA assay or other protein assay to determine the protein concentration of the cell lysate.
- Add 50 μL of cell lysate to a 96-well plate. In addition, prepare blank wells that contain 50 μL Cell Lysis Buffer. We recommend testing samples in triplicate.
- Initiate the reaction by adding 50 μL of StemTAG™ AP Activity Assay Substrate. Incubate for 10-30 minutes at 37 °C.
- Stop the reaction by adding 50 μL of 1X Stop Solution and mix by placing the plate on an orbital plate shaker for 30 seconds.
- Read the absorbance of each well at 405 nm.
- 限制
- 仅限研究用
-
- 储存条件
- 4 °C
- 储存方法
- Store all components at 4°C.
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: "A single intravenous injection of adeno-associated virus serotype-9 leads to whole body skeletal muscle transduction in dogs." in: Molecular therapy : the journal of the American Society of Gene Therapy, Vol. 16, Issue 12, pp. 1944-52, (2008) (PubMed).
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: "Anthraquinone Glycoside Aloin Induces Osteogenic Initiation of MC3T3-E1 Cells: Involvement of MAPK Mediated Wnt and Bmp Signaling." in: Biomolecules & therapeutics, Vol. 24, Issue 2, pp. 123-31, (2016) (PubMed).
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- 背景
- Embryonic stem (ES) cells are continuous proliferating stem cell lines of embryonic origin first isolated from the inner cell mass (ICM). Two distinguishing features of ES cells are their ability to be maintained indefinitely in an undifferentiated state and their potential to develop into any cell within the body. Based on previous methods developed for mouse ES cells, human ES cell lines were first established by Dr. James Thomson and colleagues. Like mouse ES cells, human ES cells express high levels of membrane alkaline phosphatase (AP) and Oct-4, a transcriptional factor critical to ICM and germline formation. However, unlike mouse ES cells, hES cells do not express stage-specific embryonic antigen (SSEA-1). In addition, prolonged propagation of hES cells is typically achieved by coculture with primary mouse embryonic fibroblasts (MEFs) serving as feeder cells. Human ES cell lines are not able to maintain their undifferentiated state in the absence of supporting feeder layer cells, even when exogenous cytokines such as leukemia inhibitory factor (LIF) and gelatin-coated plates are used.
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