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Cell Stimulation KIt(With protein transport inhibitor)

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Cell Stimulation KIt(With protein transport inhibitor)
Cat.No:
IK-CEA-1
Brand:
Solarbio
CAS号产品货号产品名称分子式分子量对应靶点产品描述
16561-29-8IP1010Phorbol 12-Myristate 13-Acetate;佛波酯C36H56O8616.83PKCPMA是一种佛波酯,是常用的 PKC 活化剂。
56092-82-1II0820Ionomycin Calcium Salt钙离子霉素C41H70O9Ca747.1calcium ionophore是一种有效的,选择性的钙离子载体 (calcium ionophore),也是一种抗生素。能够促进细胞凋亡,诱导蛋白激酶C (PKC) 激活。
20350-15-6IB0270Brefeldin A布雷非德菌素 AC16H24O4280.36protein transport是一种内酯抗生素,是蛋白质运输的特定抑制剂,可以阻断分泌蛋白和膜蛋白从内质网向高尔基体的转运。也是一种自噬 (autophagy) 和 mitophagy 抑制剂。还是一种 CRISPR/Cas9 激动剂,可抑制 HSV-1病毒,并具有抗癌活性。
22373-78-0IM1050Monensin Sodium Salt莫能霉素钠C36H61NaO11692.85protein transport是一种常用的蛋白转运抑制剂,可破坏高尔基体,抑制细胞内的蛋白转运,常用于抑制细胞因子等分泌型蛋白的分泌,使其保留在细胞内。
本套装内含1000×PMA(81μM)、Ionomycin(1.34mM)、Brefeldin A(2mM)的DMSO溶液和Monensin(10.6mM)的EtOH溶液各一支,每支溶液100μL。
在细胞内,PKC的激活可以引起下游众多蛋白激酶的磷酸化,形成级联反应,导致多种蛋白的表达,进而引起细胞的活化。PKC可以在DAG和Ca2+共同作用下被激活。PMA是一种有效的PKC激活剂;Ionomycin是一种有效的,选择性的钙离子载体。PMA常与Ionomycin联用,刺激免疫细胞活化,用于诱导体外细胞培养物产生细胞因子。

同时,也可加入蛋白转运抑制剂Monensin或Brefeldin A,抑制细胞因子外分泌,以准确分析细胞因子的表达水平。如果对分泌蛋白进行免疫检测(例如 ELISA、蛋白免疫印迹或多重蛋白检测),则无需使用Monensin或Brefeldin A对细胞进行处理。



实验案例(仅供参考

Cell

Flow Cytometric Analysis of the Percentages of Th17 Cells.(Spleen Mononuclear Cell/SMC, Thyroid Mononuclear Cell/TMC)

Phorbol 12-myristate 13-acetate (PMA, Solarbio) and Ca-ionomycin (Solarbio) were added into the 12-well plates containing the SMC or TMC suspension, and brefeldin A (BFA, Solarbio) was added after 1 h followed by incubation for 3 h (total stimulation of 4 h). Cells were then collected, washed, and surface stained with APC-labeled CD4 antibody at 4°C in the dark for 30 min. IC fixation buffer was then added followed by incubation at 4°C in the dark for 20 min. Finally, cells were washed,  resuspended in permeabilization buffer, and stained with PE-labeled IL-17A antibody at 4°C in the dark for 30 min. Flow cytometric analyses were performed using a FACScanto flow cytometer.

来源文献:Liu H, Li Y, Zhu Y, Ma L, Xue H. Notch Signaling Pathway Promotes Th17 Cell Differentiation and Participates in Thyroid Autoimmune Injury in Experimental Autoimmune Thyroiditis Mice. Mediators Inflamm. 2023 Jan 6;2023:1195149. doi: 10.1155/2023/1195149. PMID: 36643586; PMCID: PMC9839414.







Remark:
These protocols are for reference only. Solarbio does not independently validate these methods.

Note:
1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.
2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.
3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.
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