Anti-Cyclin D3/CCND3抗体[DCS2.2] (ab28283)
Key features and details
- Mouse monoclonal [DCS2.2] to Cyclin D3/CCND3
- Suitable for: Flow Cyt, WB, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
概述
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产品名称
Anti-Cyclin D3/CCND3抗体[DCS2.2]
参阅全部 Cyclin D3/CCND3 一抗 -
描述
小鼠单克隆抗体[DCS2.2] to Cyclin D3/CCND3 -
宿主
Mouse -
经测试应用
适用于: Flow Cyt, WB, IHC-Pmore details -
种属反应性
与反应: Human
预测可用于: Mouse, Rat -
免疫原
Recombinant full length protein corresponding to Human Cyclin D3/CCND3.
Sequence:MELLCCEGTRHAPRAGPDPRLLGDQRVLQSLLRLEERYVPRASYFQCVQR EIKPHMRKML AYWMLEVCEEQRCEEEVFPLAMNYLDRYLSCVPTRKAQ LQLLGAVCMLLASKLRETTPLT IEKLCIYTDHAVSPRQLRDWEVLVLG KLKWDLAAVIAHDFLAFILHRLSLPRDRQALVKK HAQTFLALCATDYT FAMYPPSMIATGSIGAAVQGLGACSMSGDELTELLAGITGTEVDCL RA CQEQIEAALRESLREASQTSSSPAPKAPRGSSSQGPSQTSTPTDVTAIHL
Database link: P30281 -
阳性对照
- WB: HeLa, K562, HAP1, HEK-293T and Jurkat cell lysates. Flow Cyt: HeLa cells. IHC-P: Human pancreas tissue.
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常规说明
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
Preservative: 0.08% Sodium azide
Constituent: PBS -
Concentration information loading...
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纯度
Protein A/G purified -
克隆
单克隆 -
克隆编号
DCS2.2 -
同种型
IgG1 -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab28283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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Flow Cyt |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 35 kDa.
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IHC-P |
Use at an assay dependent concentration.
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说明 |
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Flow Cyt
Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 35 kDa. |
IHC-P
Use at an assay dependent concentration. |
靶标
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功能
Regulatory component of the cyclin D3-CDK4 (DC) complex that phosphorylates and inhibits members of the retinoblastoma (RB) protein family including RB1 and regulates the cell-cycle during G(1)/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complex and the subsequent transcription of E2F target genes which are responsible for the progression through the G(1) phase. Hypophosphorylates RB1 in early G(1) phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also substrate for SMAD3, phosphorylating SMAD3 in a cell-cycle-dependent manner and repressing its transcriptional activity. Component of the ternary complex, cyclin D3/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex. -
序列相似性
Belongs to the cyclin family. Cyclin D subfamily.
Contains 1 cyclin N-terminal domain. -
细胞定位
Nucleus. Cytoplasm. Membrane. Cyclin D-CDK4 complexes accumulate at the nuclear membrane and are then translocated to the nucleus through interaction with KIP/CIP family members. - Information by UniProt
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数据库链接
- Entrez Gene: 896 Human
- Entrez Gene: 12445 Mouse
- Entrez Gene: 25193 Rat
- Omim: 123834 Human
- SwissProt: P30281 Human
- SwissProt: P30282 Mouse
- SwissProt: P48961 Rat
- Unigene: 534307 Human
see all -
别名
- CCND 3 antibody
- Ccnd3 antibody
- CCND3_HUMAN antibody
see all
图片
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All lanes : Anti-Cyclin D3/CCND3 antibody [DCS2.2] (ab28283) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CCND3 knockout HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
Predicted band size: 35 kDa
Observed band size: 35 kDaLanes 1-4: Merged signal (red and green). Green - ab28283 observed at 35 kDa. Red - loading control ab52901.
ab28283 Anti-Cyclin D3/CCND3 antibody [DCS2.2] was shown to specifically react with Cyclin D3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264931 (knockout cell lysate ab257876) was used. Wild-type and Cyclin D3 knockout samples were subjected to SDS-PAGE. ab28283 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Overlay histogram showing HeLa cells stained with ab28283 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab28283, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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ab28283 (4µg/ml) staining Cyclin D3/CCND3 in human pancreas, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required. -
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Cyclin D3/CCND3 (KO) knockout HAP1 whole cell lysate (20 µg)
Lane 3: HEK293 whole cell lysate (20 µg)
Lane 4: Jurkat whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab28283 observed at 33 kDa. Red - loading control, ab176560, observed at 50 kDa.ab28283 was shown to specifically recognize CCND3 in wild-type HAP1 cells along with additional cross reactive bands. No bands was observed when CCND3 knockout samples were uexamined. Wild-type and CCND3 knockout samples were subjected to SDS-PAGE. Ab28283 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Cyclin D3/CCND3 antibody [DCS2.2] (ab28283) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 35 kDa
Observed band size: 35 kDa
Additional bands at: 75 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 8 minutes
实验方案
数据表及文件
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Datasheet download
文献 (30)
ab28283 被引用在 30 文献中.
- Turco C et al. MALAT1-dependent hsa_circ_0076611 regulates translation rate in triple-negative breast cancer. Commun Biol 5:598 (2022). PubMed: 35710947
- Todorovic Z et al. Shikonin Derivatives from Onsoma visianii Decrease Expression of Phosphorylated STAT3 in Leukemia Cells and Exert Antitumor Activity. Nutrients 13:N/A (2021). PubMed: 33807148
- Xiong Y et al. Circulating Exosomal miR-20b-5p Inhibition Restores Wnt9b Signaling and Reverses Diabetes-Associated Impaired Wound Healing. Small 16:e1904044 (2020). PubMed: 31867895
- Zhang W et al. lncRNA MIAT promotes cell invasion and migration in esophageal cancer. Exp Ther Med 19:3267-3274 (2020). PubMed: 32266022
- Wu H et al. miR-138-5p suppresses glioblastoma cell viability and leads to cell cycle arrest by targeting cyclin D3. Oncol Lett 20:264 (2020). PubMed: 32989398