Caspase-2 Rabbit Recombinant mAb

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  • WB
规格 价格 库存 购买数量
20ul RMB 447.02 现货
100ul RMB 1500.19 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) 48kDa
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 Caspase-2 Rabbit Recombinant mAb可检测Caspase-2内源性水平。
背景 Caspases是半胱氨酸蛋白酶,主要有两种功能:促炎症细胞因子的加工和激活、以及凋亡过程中多种蛋白的剪接。在人类细胞中表达的caspases中,caspases 1、4、5主要参与细胞因子加工,而caspases 3、6、7、8、9、10主要参与凋亡过程的蛋白剪接。Caspases 3、8、14在细胞增殖和分化中也发挥了作用。在內源性和外源性细胞死亡信号中,Caspase 2被激活,它是一种下游caspase。Caspase-2定位在核内,参与DNA损伤反应、细胞周期调节和肿瘤抑制。

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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