人CCND1 (Cyclin D1) knockout HeLa cell裂解物(ab263808)
概述
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产品名称
人CCND1 (Cyclin D1) knockout HeLa cell裂解物
参阅全部 Cyclin D1 试剂盒 -
产品概述
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 1. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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说明
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
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经测试应用
适用于: WBmore details
性能
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存放说明
Store at -80°C. Please refer to protocols. -
组件 1 kit ab255457 - Human CCND1 knockout HeLa cell lysate 1 x 100µg ab255552 - Human wild-type HeLa cell lysate 1 x 100µg -
研究领域
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8, 12 CSF1PO: 9, 10
靶标
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功能
Essential for the control of the cell cycle at the G1/S (start) transition. -
疾病相关
Note=A chromosomal aberration involving CCND1 may be a cause of B-lymphocytic malignancy, particularly mantle-cell lymphoma (MCL). Translocation t(11;14)(q13;q32) with immunoglobulin gene regions. Activation of CCND1 may be oncogenic by directly altering progression through the cell cycle.
Note=A chromosomal aberration involving CCND1 may be a cause of parathyroid adenomas. Translocation t(11;11)(q13;p15) with the parathyroid hormone (PTH) enhancer.
Defects in CCND1 are a cause of multiple myeloma (MM) [MIM:254500]. MM is a malignant tumor of plasma cells usually arising in the bone marrow and characterized by diffuse involvement of the skeletal system, hyperglobulinemia, Bence-Jones proteinuria and anemia. Complications of multiple myeloma are bone pain, hypercalcemia, renal failure and spinal cord compression. The aberrant antibodies that are produced lead to impaired humoral immunity and patients have a high prevalence of infection. Amyloidosis may develop in some patients. Multiple myeloma is part of a spectrum of diseases ranging from monoclonal gammopathy of unknown significance (MGUS) to plasma cell leukemia. Note=A chromosomal aberration involving CCND1 is found in multiple myeloma. Translocation t(11;14)(q13;q32) with the IgH locus. -
序列相似性
Belongs to the cyclin family. Cyclin D subfamily. -
翻译后修饰
Phosphorylation at Thr-286 by MAP kinases is required for ubiquitination and degradation following DNA damage. It probably plays an essential role for recognition by the FBXO31 component of SCF (SKP1-cullin-F-box) protein ligase complex.
Ubiquitinated, primarily as 'Lys-48'-linked polyubiquitination. Ubiquitinated by a SCF (SKP1-CUL1-F-box protein) ubiquitin-protein ligase complex containing FBXO4 and CRYAB (By similarity). Following DNA damage it is ubiquitinated by some SCF (SKP1-cullin-F-box) protein ligase complex containing FBXO31. Ubiquitination leads to its degradation and G1 arrest. Deubiquitinated by USP2; leading to stabilize it. -
细胞定位
Nucleus. - Information by UniProt
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别名
- AI327039
- B cell CLL/lymphoma 1
- B cell leukemia 1
see all
相关产品
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KO cell lines
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab263808于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB |
Use at an assay dependent concentration.
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说明 |
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WB
Use at an assay dependent concentration. |
图片
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Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: CCND1 knockout HeLa cell lysate (20µg)Lanes 1- 2: Merged signal (red and green). Green - ab40754 observed at 36 kDa. Red - loading control ab130007 observed at 124 kDa.
ab40754 Recombinant Anti-Cyclin D1 antibody [EP272Y] was shown to specifically react with CCND1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255348 (knockout cell lysate ab263808) was used. Wild-type and CCND1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab40754 and Anti-Vinculin antibody [VIN-54] were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: CCND1 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab16663 observed at 36 kDa. Red - loading control ab130007 observed at 124 kDa.
ab16663 Recombinant Anti-Cyclin D1 antibody [SP4] was shown to specifically react with CCND1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255348 (knockout cell lysate ab263808) was used. Wild-type and CCND1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab16663 and Anti-Vinculin antibody [VIN-54] were incubated overnight at 4°C at 1 in 200 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging. -
Homozygous: 5 bp deletion in exon 1
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
文献 (0)
ab263808 尚未被引用在任何文献中。