Cyclin D1 Rabbit Recombinant mAb

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  • WB
规格 价格 库存 购买数量
20ul RMB 447.58 现货
100ul RMB 1500.47 现货
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Cited by 5 Publications

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) 36kDa
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 Cyclin D1 Rabbit Recombinant mAb可检测cyclin D1的内源性水平。
背景 Cyclin D1在G1期早到中期,是细胞外信号的关键感应和集合分子。通过结合CDKs和组蛋白乙酰基转移酶、组蛋白脱乙酰基酶发挥其功能,调节细胞增殖、分化相关基因的染色体结构。Cyclin D1是全酶的调节亚基,磷酸化pRb,同时被cyclinE/CDK2连续磷酸化,使pRb失活。pRb是G1期的gatekeeper,通过这一限制点将引起DNA合成。在细胞粘附、迁移和原代骨髓巨噬细胞的指导迁移中,cyclin D1也是必需的。Cyclin D1与转录因子或共激活剂包括HATs和HDACs相结合,调节转录和表观遗传变化。

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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