ATF2 Rabbit Recombinant mAb

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  • WB
规格 价格 库存 购买数量
20ul RMB 447.49 现货
100ul RMB 1500.76 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
MW (kDa) 70kDa
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 ATF2 Rabbit Recombinant mAb可检测ATF2的内源性水平。
背景 Activating transcription factor 2 (ATF2)在黑素瘤中具有致癌活性,在非恶性皮肤肿瘤和乳腺癌中具有肿瘤抑制活性。ATF2被JNK、p38、ERK1磷酸化,成为具有转录活性的蛋白。在受到亚基和细胞因子刺激后,ATF2对低氧、渗透应力、DNA损伤、病毒感染和细胞死亡等细胞过程中发挥作用。ATF2的转录作用取决于其二聚体搭档,其搭档一般是AP-1家族成员。ATF2同源二聚体的转录活性差,所以其异源二聚体对于其发挥转录功能很重要。根据不同的二聚体搭档,ATF2结合靶基因上不同的相应元件,从而启动不同的转录程序。在未受刺激的情况下,ATF2的C端DNA结合区域和其N端激活区域发生分子内结合,使其处于转录失活形式。当转录辅激活子、组蛋白乙酰基转移酶CBP或腺病毒13S E1A与bZIP区域结合时,减轻分子内抑制作用,激活转录。ATF2还可与p300互相作用。在小鼠胚胎性癌细胞中,复合体的形成对于在细胞分化的诱导剂刺激下c-Jun基因的表达调节很重要。当ATF2与ATP依赖性的解旋酶TIP49b相互作用时,将下调ATF2的转录活性。

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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