| SKU-Pack Size | Availability | Price | Quantity |
| abs955-50tests | In stock | $420.00 | - + |
Request bulk quotation| The name of the reagent | volume | Preservation conditions |
| Lysis buffer | 30ml | 4 ℃ |
| 10*Wash buffer | 50ml | 4 ℃ |
| 1*SDS Sample buffer | 1ml | 4 ℃ |
| Protein A Agarose beads | 250ul | 4 ℃ |
| Protein GAgarose beads | 250ul | 4 ℃ |
2.use method
A. Prepare cell lysates
1. Blot the medium dry. A fresh medium containing regulatory molecules was added to allow the cells to be treated within a predetermined time.
2. Cells were collected and washed once with 1 X PBS precooled with ice after removing the culture medium.
3. Remove PBS and add 0.5 mL ice precooled Lysis buffer to each cell plate (10 cm) and incubate on ice for at least 5 minutes.
4. Scrape the cells from the plate and transfer the extract to a microcentrifuge tube. On ice.
5. Perform ultrasonic crushing on the ice for 3 times, 5 seconds each time.
6. Microcentrifugation at 4 °C at 14,000 x g for 10 min. Transfer supernatant to new tube. The supernatant is the cell lysate. If necessary, the pyrolysis may be stored at -80 °C.
Note: After the cell lysates are prepared, western blotting can be performed to determine the presence of the target protein in the cell lysates.
B. Immunoprecipitation method
A) Cell lysate pre-clearance (optional step)
1. Add 5 μ L Protein A and 5 μ L Protein G to 500 μ L (containing 200-1000 ug total Protein) cell lysate.
2. Rotate and incubate at 4 °C for 30-60 minutes.
3. Centrifugation at 12000g at 4°C for 1 min. Reserve supernatant.
B) antigen antibody binding
1. Add 500 μ L (200 -- 1000 ug total protein) cell lysate (can be pre-cleaned cell lysate) to the new centrifuge tube.
2. Add monoclonal/polyclonal antibody (1-5 μg) to target protein.
3. Homologous antibodies of non-specific immunity were used as control.
4. Blend gently at 4 °C overnight.
C) Precipitation of immune complexes
1. After incubation overnight, add 5 μ L Protein A and 5 μ L Protein G.
2. Mix gently at 4 °C for 1-3 hours or overnight.
3. Centrifuge at 12,000 g for 1 minute, retain the precipitation.
4. Use 0.5 ml 1*Wash buffer to clean the precipitation, centrifuge at 12,000 g for 1 minute, and retain the precipitation.
5. Repeat step 4 for a total of 3 times.
Note: When cleaning and removing the supernatant, care should be taken to avoid suction of the filler
C. Western blotting was used for analysis
1. Suspend the precipitate in 20-40 μ L 1*SDS sample buffer, swirl the precipitate, then centrifuge for 30 seconds to remove beads and liquid from tube wall to tube bottom.
2. Heat the sample to 95-100 °C for 2-5 minutes, then centrifuge at 14,000 g for 1 minute, remove supernatant.
3. Load sample (15 -- 30 μ L) onto SDS-PAGE gel.
4. Samples were analyzed by western blotting.
3. Method of kit preservation
The kit is stored at 4℃ and effective for 1 year.
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Unsuccessful submission? Please contact info@absin.cn.
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Journal of Oncology. 2022 Jan 11 .
effect： 4.375
- Sodium-Glucose Co-transporter-2 Inhibitor of Dapagliflozin Attenuates Myocardial Ischemia/Reperfusion Injury by Limiting NLRP3 Inflammasome Activation and Modulating Autophagy
Yong-Wei Yu, Jia-Qun Que, Shuai Liu, Kai-Yu Huang, Lu Qian, Ying-Bei Weng, Fang-Ning Rong, Lei Wang, Ying-Ying Zhou, Yang-Jing Xue, Kang-Ting Ji
Frontiers in Cardiovascular Medicine. 2022 Jan 10 .
effect： 4.79
- EGLN1 induces tumorigenesis and radioresistance in nasopharyngeal carcinoma by promoting ubiquitination of p53 in a hydroxylase-dependent manner
Lu Sun, Cheng Wu, Jun Ming, Ergang Guo, Wei Zhang, Lingling Li, Guoqing Hu
Journal of Cancer. 2022 Jan 10 ; 7
effect： 4.207
- Sodium-Glucose Co-transporter-2 Inhibitor of Dapagliflozin Attenuates Myocardial Ischemia/Reperfusion Injury by Limiting NLRP3 Inflammasome Activation and Modulating Autophagy
Yong-Wei Yu, Jia-Qun Que, Shuai Liu, Kai-Yu Huang, Lu Qian, Ying-Bei Weng, Fang-Ning Rong, Lei Wang, Ying-Ying Zhou, Yang-Jing Xue, Kang-Ting Ji
Frontiers in Cardiovascular Medicine. 2022 Jan 10 .
effect： 6.050
- Harnessing chlorin e6 loaded by functionalized iron oxide nanoparticles linked with glucose for target photodynamic therapy and improving of the immunogenicity of lung cancer
Ting-Ting Yu, Xing-Chun Peng, Mei-Fang Wang, Ning Han, Hua-Zhen Xu, Qi-Rui Li, Liu-Gen Li, Xiang Xu, Qian-Li Ma, Bin Liu, Jue Wang, Li Zhao, Xiao Chen, Tong-Fei Li
Journal of Cancer Research and Clinical Oncology. 2022 Jan 08 .
effect： 4.553
- RNA-binding motif protein 10 represses tumor progression through the Wnt/β- catenin pathway in lung adenocarcinoma
Yingyue Cao, Jianxiong Geng, Xin Wang, Qingwei Meng, Shanqi Xu, Yaoguo Lang, Yongxu Zhou, Lishuang Qi, Zijie Wang, Zixin Wei, Yan Yu, Shi Jin, Bo Pan
International Journal of Biological Sciences. 2022 Jan 01 ; 1
effect： 6.58
- 基于NLRP1炎症小体通路研究自噬在金丝桃苷抗小鼠心肌梗死中的作用及机制研究
杨永康
安徽医科大学. 2021/03/01 00:00:00 undefined undefined .
- Cancer-Associated Fibroblasts Promote Vascular Invasion of Hepatocellular Carcinoma via Downregulating Decorin-integrin β1 Signaling
Xiaobo Zheng, Peng Wang, Li Li, Jing Yu, Chune Yu, Liangliang Xu, Lian Li, Fuzhen Dai, Lei Feng, Hong Zou, Xiaobo Chen6, Ming Zhang, Mingqing Xu
Frontiers in Cell and Developmental Biolog. 2021 Aug 24 .
effect： 5.186
- Mitochondria damage in ambient particulate matter induced cardiotoxicity: Roles of PPAR alpha/PGC-1 alpha signaling
Qixiao Jianga, Andong Jia, Daochuan Lib, Limei Shia, Mengyu Gaoa, Na Lvc, Ying Zhanga, Rong Zhangd, Rui Chene, Wen Chenb, Yuxin Zhenga, Lianhua Cui
Environmental Pollution. 2021 Aug 14 .
effect： 8.071
- Mitochondria damage in ambient particulate matter induced cardiotoxicity: Roles of PPAR alpha/PGC-1 alpha signaling
Qixiao Jianga, Andong Jia, Daochuan Lib, Limei Shia, Mengyu Gaoa, Na Lvc, Ying Zhanga, Rong Zhangd, Rui Chene, Wen Chenb, Yuxin Zhenga, Lianhua Cui
Environmental Pollution. 2021 Aug 14 .
effect： 8.071
- Pannexin-1 promotes the invasion of pituitary adenoma by modulating ATP release
Xiaoshuang Tang
Research Square. 2021 Aug 12 .
- The role and mechanism of hyperoside against myocardial infarction in mice by regulating autophagy via NLRP1 inflammation pathway
YongkangYang,JingLi,TingcaiRao,ZhiruiFang,JunyanZhang
Journal of Ethnopharmacology. 2021 Aug 10 .
effect： 4.360
- Porcine beta-defensin 2 provides protection against bacterial infection by a direct bactericidal activity and alleviates inflammation via interference with the TLR4/NF-κB Pathway
Chao Huang, Xi Yang, Jing Huang, Xiao Liu, Xiaoyu Yang,Hui Jin, Qi Huang,Lu Li,Rui Zhou
Frontiers in Immunology. 2021 Jul 18 .
effect： 7.561
- Role and mechanism of chaperones calreticulin and ERP57 in restoring trafficking to mutant HERG‑A561V protein
Yujia Wu, Xiaoyan Huang, Zequn Zheng, Xi Yang, Yanna Ba, Jiangfang Lian
International Journal of Molecular Medicine. 2021 Jul 01 .
effect： 2.943
- 枸杞多糖促进小肠内分泌细胞分泌肠促胰液素
邓姝颖; 白莉莉; 薛阳; 陈康; 蔡慧珍
宁夏医科大学学报. 2021 Jun 30 .
- Upregulation of SGOL2 Predicts Poor Prognosis and Facilitates Hepatocellular Carcinoma Progression via Dysregulating the Cell Cycle by Directly Activating MAD2
Qingqing Hu, Xiaochu Hu, Yalei Zhao, Lingjian Zhang, Ya Yang, lanjuan li
Research Square. 2021 Jun 08 .
- LncRNA MIR99AHG/miR-136-5p/USP4/ACE2信号轴调控肺纤维化及其机制研究
李会
武汉科技大学. 2021 May 01 .
- The role of E26 transformation-specific variant transcription factor 5 in colorectal cancer cell proliferation and cell cycle progression
Yi Peng, Haoran Feng, Changgang Wang, Zijia Song, Yaqi Zhang, Kun Liu, Xi Cheng, Ren Zhao
Cell Death & Disease. 2021 Apr 30 .
effect： 5.989
- YY1-modulated long non-coding RNA SNHG12 promotes gastric cancer metastasis by activating the miR-218-5p/YWHAZ axis
Tianqi Zhang, Maneesh Kumarsing Beeharry, Zhenqiang Wang, Zhenggang Zhu, Jianfang Li, Chen Li
International Journal of Biological Sciences. 2021 Apr 12 ; 7
effect： 4.615
- USP15 participates in DBP-induced testicular oxidative stress injury through regulating the Keap1/Nrf2 signaling pathway
Lei Zhang, Xian Gao, Zhiqiang Qin, Xiaokai Shi, Kai Xu, Shangqian Wang, Min Tang, Wei Wang, Shengling Gao, Li Zuo, Lifeng Zhang, Wei Zhang
Science of The Total Environment. 2021 Apr 09 .
effect： 6.223
- Caspofungin suppresses brain cell necroptosis in the ischemic stroke rat via up-regulation of Pellino3
Yi-Yue Zhang, Jing Tian, Zi-Mei Peng, Ya-Wei Peng, Xiao-Jie Zhang, Zhong-Yang Hu, Xiu-Ju Luo, Jun Peng
Research Square. 2021 Mar 30 .
- MYCT1通过RACK1介导的酶选择性翻译调控糖原分流的机制研究
丁东雪
中国医科大学. 2021 Feb 01 .
- Ubiquitin-specific protease 7 promotes ferroptosis via activation of the p53/TfR1 pathway in the rat hearts after ischemia/reperfusion
Li-Jing Tang, Yuan-Jing Zhou, Xiao-Ming Xiong, Nian-Sheng Li, Jie-Jie Zhang, Xiu-Ju Luo, Jun Peng
Free Radical Biology and Medicine. 2021 Jan 01 .
effect： 7.376
- Comprehensive analysis of ubiquitin‐specific protease 1 reveals its importance in hepatocellular carcinoma
Yalei Zhao, Chen Xue, Zhongyang Xie, Xiaoxi Ouyang, Lanjuan Li
Cell Prolif. 2020 Sep 19 .
effect： 5.753
- YY1-modulated Long Non-coding RNA SNHG12 Predicts and Promotes Gastric Cancer Metastasis via Activating miR-218-5p/YWHAZ-dependent βcatenin: A Bed to Bench Approach
Tian Qi Zhang, Qingqiang Dai, Maneesh Kumarsing Beeharry, Zhenqiang Wang, Liping Su, Chen Li
Research Suare. 2020 Aug 24 .
- SNW1 interacts with IKKγ to positively regulate antiviral innate immune responses against influenza A virus infection
Qiao Zhang, Taizhen Liang, Shuyin Gu, Yilu Ye, Shuwen Liu
Microbes and Infection. 2020 Aug 14 .
effect： 2.373
- Fibrinogen-like protein 2 aggravates nonalcoholic steatohepatitis via interaction with TLR4, eliciting inflammation in macrophages and inducing hepatic lipid metabolism disorder
Junjian Hu,Hongwu Wang,Xitang Li,Yonggang Liu,Yuqiang Mi,Hongyan Kong,Dong Xi,Weiming Yan,Xiaoping Luo,Qin Ning, Xiaojing Wang
Theranostics. 2020 Aug 01 ; 21
effect： 8.579
- MicroRNA-146a在阿霉素诱导心肌损伤中的保护作用及机制研究
潘建安
上海交通大学. 2020 Jun 04 .
- AZIN-1小分子抑制剂抗非小细胞肺癌A549活性的研究
王顺超
三峡大学. 2020 May 01 .
- CNPase在后发性白内障上皮-间质转化中的作用和机制研究
李玥
天津医科大学. 2020 May 01 .
- PP4R1 interacts with HMGA2 to promote non‐small‐cell lung cancer migration and metastasis via activating MAPK/ERK‐induced epithelial‐mesenchymal transition
Bin Wang, Lin‐Yue Pan, Ning Kang, Xiao‐Yong Shen
Mol Carcinog. 2020 Feb 19 ; 5
effect： 3.411
- Berberine increases glucose uptake and intracellular ROS levels by promoting Sirtuin 3 ubiquitination
Wei Li, Dan Li, Hongying Kuang, Xiaoling Feng, Wenxia Ai, Ying Wang, Simao Shi, Jing Chen, Ruifeng Fan
BIOMED PHARMACOTHER. 2020 Jan 01 .
effect： 3.743
- Red blood cell–derived nanoerythrosome for antigen delivery with enhanced cancer immunotherapy
XIAO HAN,SHUFANG SHEN,QIN FAN,GUOJUN CHEN,EDIKAN ARCHIBONG,GIANPIETRO DOTTI,ZHUANG LIU,ZHEN GU, CHAO WANG
science advances. 2019 Aug 23 .
effect： 14.136
- DUSP8对MSI-H型和MSS型人结直肠癌细胞生长和凋亡的影响及机制研究
丁涛
遵义医科大学. 2019 May 01 .
- MMP9通过结合IFNAR1抑制IFN/JAK/STAT通路促进EV71复制
蒋曦
北京协和医学院. 2019 May 01 .
- Tomatidine通过调控破骨细胞分化减少雌激素缺乏导致的骨量丢失的实验研究
胡斌
浙江大学. 2019 May 01 ; 3
- 胰岛再生源蛋白RegⅢγ对大鼠肝细胞BRL-3A生长的作用及机制研究
赵聪聪
河南师范大学. 2018 May 01 ; 1
- Rapamycin attenuated podocyte apoptosis via upregulation of nestin in Ang II-induced podocyte injury
Huimin Shi, Yajie Zhao, Tiantian He, Xianli Wen, Gaoting Qu, Shanwen Li, Weihua Gan, Aiqing Zhang
Research Square.
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