Anti-Cdk6 抗体 [EPR4515] - BSA and Azide free (ab222395)
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
- 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] - BSA and Azide free
Cdk6 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR4515] to Cdk6 - BSA and Azide free -
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: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details
適用なし: IP -
種交差性
交差種: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: Jurkat, K562, HeLa, HAP1, and 293T cell lysates. IHC-P:Human tonsil tissue. ICC/IF: HeLa and HAP1 cells.
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特記事項
ab222395 is the carrier-free version of ab124821.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
バッファー
pH: 7.2
Constituent: PBS -
キャリア・フリー
はい -
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
- Anti-Cdk6 antibody [EPR4515] (ab124821)
- Alexa Fluor® 488 Anti-Cdk6 antibody [EPR4515] (ab198944)
- Alexa Fluor® 647 Anti-Cdk6 antibody [EPR4515] (ab198946)
- PE Anti-Cdk6 antibody [EPR4515] (ab305713)
- APC Anti-Cdk6 antibody [EPR4515] (ab305714)
- HRP Anti-Cdk6 antibody [EPR4515] (ab305715)
- Alexa Fluor® 594 Anti-Cdk6 antibody [EPR4515] (ab310613)
- Alexa Fluor® 555 Anti-Cdk6 antibody [EPR4515] (ab312141)
- Alexa Fluor® 568 Anti-Cdk6 antibody [EPR4515] (ab312627)
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Compatible Secondaries
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Conjugation kits
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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ab222395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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (1) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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特記事項 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab124821).
Lanes 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124821).
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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. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124821).
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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. -
ab124821 at 1/100 dilution, staining Cdk6 in formalin-fixed paraffin-embedded human tonsil tissue by immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124821).
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Clone EPR4515 (ab222395) has been successfully conjugated by Abcam. This image was generated using Anti-Cdk6 antibody [EPR4515] (Alexa Fluor® 647). Please refer to ab198946 for protocol details.
ab198946 staining Cdk6 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), 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 overnight at +4°C with ab198946 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).
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Clone EPR4515 (ab222395) has been successfully conjugated by Abcam. This image was generated using Anti-Cdk6 antibody [EPR4515] (Alexa Fluor® 488). Please refer to ab198944 for protocol details.
ab198944 staining Cdk6 in HeLa cells. The cells were fixed with 100% methanol (5min), 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 overnight at +4°C with ab198944 at 1/100 dilution (shown in green) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml. This was followed by an incubation at room temperature for 1h with ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed, at 1µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
プロトコール
データシートおよび資料
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Datasheet download
Certificate of Compliance
参考文献 (6)
ab222395 は 6 報の論文で使用されています。
- Li T et al. Massive renal urothelial carcinoma with renal vein tumor thrombus, pancreatic infiltration and adrenal metastasis: A case report. Oncol Lett 12:1129-1131 (2016). IHC-P ; Human . PubMed: 27446406
- Wang G et al. Simvastatin induces cell cycle arrest and inhibits proliferation of bladder cancer cells via PPAR? signalling pathway. Sci Rep 6:35783 (2016). WB . PubMed: 27779188
- Yao CJ et al. Activation of p53/miR-34a Tumor Suppressor Axis by Chinese Herbal Formula JP-1 in A549 Lung Adenocarcinoma Cells. Evid Based Complement Alternat Med 2016:5989681 (2016). WB ; Human . PubMed: 28074102
- He B et al. Upregulated microRNA-143 inhibits cell proliferation in human nasopharyngeal carcinoma. Oncol Lett 12:5023-5028 (2016). WB ; Human . PubMed: 28105209
- Wang Y et al. Downregulation of microRNA-33a promotes cyclin-dependent kinase 6, cyclin D1 and PIM1 expression and gastric cancer cell proliferation. Mol Med Rep 12:6491-6500 (2015). WB ; Human . PubMed: 26352175
- Chen MT et al. ZFP36L1 promotes monocyte/macrophage differentiation by repressing CDK6. Sci Rep 5:16229 (2015). WB ; Human . PubMed: 26542173