p38 MAPK Rabbit Recombinant mAb

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  • WB
规格 价格 库存 购买数量
20ul RMB 447.73 现货
100ul RMB 1500.29 现货
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400-668-6834

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Cited by 1 Publication

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) 41kDa
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 p38 MAPK Rabbit Recombinant mAb可检测p38 MAPK的内源性水平。
背景 P38 MAPK是丝裂原激活蛋白激酶类蛋白,对压力刺激如细胞因子、紫外线照射、热激和渗透压休克产生响应,参与细胞分化、凋亡和自噬。目前发现的有4种p38 MAPKs:p38-α (MAPK14), -β (MAPK11), -γ (MAPK12 / ERK6)和-δ (MAPK13 / SAPK4)。与ERK信号通路不同,哺乳类的p38 MAPK家族成员(alpha, beta, gamma和delta),在JNK参与或不参与的情况下,主要调节炎症和应激反应。

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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