TNF Receptor II Rabbit Recombinant mAb

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  • WB
规格 价格 库存 购买数量
20ul 447.97 现货
100ul 1500.02 现货
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400-668-6834

info@selleck.cn

 

使用信息

抗体应用 WB,ELISA
稀释比例
WB
1:1000
反应性 Human Mouse Rat
MW (kDa) 73kDa
抗体类型 Rabbit
浓度 1mg/ml
储存液配方 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide.
储存条件
(自收到货起)
Store at –20°C.

Datasheet & SDS

生物描述

特异性 TNF Receptor II Rabbit Recombinant mAb可检测TNFR2的内源性水平。
背景 肿瘤坏死因子(TNF)是一种多效性细胞因子,参与调节多种生理功能:细胞生长、炎症反应、肿瘤生成、病毒复制、感染性休克和自身免疫反应。TNF通过两种受体发挥作用:TNFR1和TNFR2。TNFR1和TNFR2具有不同的表达模式。TNFR1在淋巴系统、几乎所有体内细胞中广泛表达,发挥TNF广泛功能。TNFR2的定位则更为精确,位于淋巴细胞的某些特定种群中:包括Tregs、内皮细胞、小神经胶质、神经元亚型、少突细胞、心肌细胞、胸腺细胞、胰岛、人间充质干细胞。尽管TNFR1和TNFR2的功能有所重叠,但依据细胞的激活状态和其他因子的相互作用,TNFR1主要介导凋亡反应,而TNFR2介导细胞生存相关反应。相似地,尽管TNFR1和TNFR2的胞内信号途径有所重叠和交叉,但本质上是不同的胞内信号途径。TNFR2信号依赖于TRAF2及NF-κB的激活和入核。

实验方法

WB

Western Blotting

Sample preparation

1. Aspirate media from cultures and Wash the cells with 1X PBS.
2. Lyse cells by adding 1X SDS sample buffer and transfer the extract to a microcentrifuge tube. Keep onice.
3. Sonicate for 10–15 sec to complete cell lysis and shear DNA.
4. Heat a 20 µl sample to 95–100°C for 5 min, then cool on ice.
5. Centrifuge for 5 min (with Microcentrifuge).
6. Load appropriate volumes of samples onto SDS-PAGE gel (loading quantity of protein sample depends on the concentration of extracted proteins).
NOTE: At the same time, please load the pre-stained molecular weight markers to determine molecular weights and verify electrotransfer.
7. Electrotransfer to nitrocellulose/PVDF membrane.

Membrane Blocking and Antibody Incubations

a. Blocking

1. (Optional) After transfer, wash the transferred membrane with TBS for 5 min at room temperature.
2. Incubate the membrane in the blocking buffer for 1 hr at room temperature.
3. Wash three times for 5 min each with TBST.

b. Antibodies Incubation

1. Incubate membrane and primary antibody (at the appropriate dilution and diluent recommended) in a primary antibody dilution buffer with gentle agitation overnight at 4°C.
2. Wash three times for 5 min each with TBST.
3. Incubate membrane with an appropriate second antibodydissolved in the blocking buffer with gentle agitation for 1 hr at room temperature.
4. Wash three times for 5 min each with TBST.
5. Proceed with detection.

Detection of Proteins

1. After antibodies incubation, Wash membrane three times for 5 minutes in TBST.
2. PrepareECL Reagent (or other chromogenic agents/substrate according to your second antibody). Mix well.
3. Incubate substrate with membrane for 1 minute, remove excess solution (membrane remains wet), wrap in plastic and expose to X-ray film.

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