Anti-Cdk6 抗体 [EPR4515] (ab124821)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4515] to Cdk6
- Suitable for: ICC/IF, WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Cdk6 antibody [EPR4515]
Cdk6 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR4515] to Cdk6 -
由来種
Rabbit -
特異性
Based on the immunogen sequence, we do not expect the antibody to cross-react with other CDK family members. No cross-reactivity testing has been performed.
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アプリケーション
適用あり: ICC/IF, WB, IHC-P, Flow Cyt (Intra)more details
適用なし: IP -
種交差性
交差種: Human -
免疫原
Synthetic peptide. within Human Cdk6 aa 300 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: Q00534 -
ポジティブ・コントロール
- WB: Jurkat, K562, HAP1, HeLa and 293T cell lysates. IHC-P: Human tonsil tissue. ICC/IF: HeLa and wild-type HAP1 cells.
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特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR4515 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab124821の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ICC/IF |
1/500.
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WB | (1) |
1/50000 - 1/200000. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa).
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
特記事項 |
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ICC/IF
1/500. |
WB
1/50000 - 1/200000. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ターゲット情報
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機能
Probably involved in the control of the cell cycle. Interacts with D-type G1 cyclins. -
配列類似性
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
Contains 1 protein kinase domain. - Information by UniProt
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参照データベース
- Entrez Gene: 1021 Human
- Omim: 603368 Human
- SwissProt: Q00534 Human
- Unigene: 119882 Human
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別名
- CDK 6 antibody
- CDK6 antibody
- CDK6_HUMAN antibody
see all
画像
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All lanes : Anti-Cdk6 antibody [EPR4515] (ab124821) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CDK6 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 37 kDaLanes 1- 2: Merged signal (red and green). Green - ab124821 observed at 37 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab124821 was shown to react with Cdk6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab266059 (knockout cell lysate ab257088) was used. Wild-type HeLa and CDK6 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab124821 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 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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ab124821 staining Cdk6 in wild-type HAP1 cells (top panel) and Cdk6 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab124821 at 1/500 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
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ab124821 at 1/100 dilution, staining Cdk6 in Formalin fixed Paraffin-embedded Human tonsil tissue by Immunohistochemistry.
Heat mediated antigen retrieval was performed with 0.01M citrate buffer, pH 6.0.
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Lanes 1-2 : Anti-Cdk6 antibody [EPR4515] (ab124821) at 1/10000 dilution
Lanes 3-4 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution
Lanes 1 & 3 & 5 : Wild-type HAP1 cell lysate
Lanes 2 & 4 & 6 : CDK6 knockout HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 37 kDaLanes 1 and 2: Green signal from target - ab124821 observed at 37 kDa
Lanes 3 and 4: Red signal from loading control - ab8226 observed at 42 kDa
Lanes 5 and 6: Merged (red and green) signal
ab124821 was shown to specifically react with CDK6 when CDK6 knockout samples were used. Wild-type and CDK6 knockout samples were subjected to SDS-PAGE. ab124821 and ab8226 (loading control to beta actin) were diluted at 1/10 000 and 1/1000 respectively and incubated overnight at 4°C. 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 1/10,000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-Cdk6 antibody [EPR4515] (ab124821) at 1/1000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : K562 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : 293T cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat-anti-rabbit HRP at 1/2000 dilution
Predicted band size: 37 kDa
Observed band size: 37 kDa -
ICC/IF image of ab124821 at 1/100 dilution, staining Cdk6 in HeLa cells.
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Overlay histogram showing HeLa cells stained with ab124821 (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 (ab124821, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (91)
ab124821 は 91 報の論文で使用されています。
- Kitamura S et al. Cyclin-dependent kinase 4/6 inhibitors suppress tumor growth in extramammary Paget's disease. Cancer Sci 113:802-807 (2022). PubMed: 34866279
- Liu Y et al. Activation of miR-500a-3p/CDK6 axis suppresses aerobic glycolysis and colorectal cancer progression. J Transl Med 20:106 (2022). PubMed: 35241106
- Shi M et al. Cytokeratin 13 promotes radiotherapy sensitivity of nasopharyngeal carcinoma by downregulating the MEK/ERK pathway. IUBMB Life 74:543-553 (2022). PubMed: 35426961
- Sun R et al. CDK6 Immunophenotype Implicates Potential Therapeutic Application of CDK4/6 Inhibitors in Urothelial Carcinoma. Front Oncol 12:819003 (2022). PubMed: 35463324
- Xu Y et al. Knockdown of lncRNA ENST00000603829 Inhibits the Proliferation and Invasion of Salivary Gland Pleomorphic Adenoma through Regulating Cyclin D1. Appl Bionics Biomech 2022:8805305 (2022). PubMed: 35528529