用户验证的应用
经验证应用
| 项目 | 详情 |
|---|---|
| WB | Western blot analysis of extracts from Hela(lane 2(20μg), K-562(lane 3(20μg) and HEK293(lane 4(20μg) using Cdk6 Antibody( HY-P80079 ) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature |
| IHC-P | Immunohistochemical analysis of paraffin-embedded human colon cancer using CDK6 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80079, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody ( HY-P83652 ). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human ovarian cancer using CDK6 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80079, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody ( HY-P83652 ). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human bladder cancer using CDK6 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80079, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody ( HY-P83652 ). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human liver using CDK6 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80079, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody ( HY-P83652 ). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human stomach cancer using CDK6 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80079, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody ( HY-P83652 ). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human prostate cancer using CDK6 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80079, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody ( HY-P83652 ). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human thyroid cancer using CDK6 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80079, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody ( HY-P83652 ). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human tonsil using CDK6 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80079, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody ( HY-P83652 ). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. Immunohistochemical analysis of paraffin-embedded human kidney using CDK6 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH 9.0) for 14 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with TBST, and then probed with the primary antibody (HY-P80079, 1/200) for 30 minutes at room temperature. The detection was performed using Polymer HRP-conjugated Goat Anti-Mouse/Rabbit lgG(H&L) secondary antibody ( HY-P83652 ). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
| FC | Flow cytometric analysis of 1X10^6 Hela cells labeling Cdk6 Antibody(HY-P80079, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/50 dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L ( HY-P8002 ) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black). |
| ICC | Immunocytochemistry analysis of HepG2 cells labeling Cdk6 with Cdk6 Antibody ( HY-P80079 ) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Cdk6 Antibody ( HY-P80079 ) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue). Immunocytochemistry analysis of Hela cells labeling Cdk6 with Cdk6 Antibody ( HY-P80079 ) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Cdk6 Antibody ( HY-P80079 )at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue) |
推荐稀释比例
- WB1:500-1:2000
- ICC/IF1:50-1:200
- IF-Tissue1:50-1:200
- IHC-P1:50-1:200
- FC1:50-1:100
详细信息
免疫原Synthetic peptide corresponding to Human Cdk6.AA range:277-326.
宿主/亚型Rabbit / IgG
抗体类型Recombinant,Monoclonal
全称CDK6 抗体 (YA511)
理论分子量37 kDa
实际/观测分子量37 kDa 请注意：因蛋白存在修饰或聚体等情况，以实测为准，预测仅为参考。
产品形式溶液
偶联物Non-conjugated
浓度不同批次浓度可能有所不同，具体浓度请参见对应批次的COA。 批次浓度查询
储存缓冲液Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.
储存条件保存条件 & 期限: Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.
运输条件: Shipping with blue ice.
纯化方式Protein A affinity purified.
Gene ID1021 [ NCBI ]
RRIDAB_3102292
UniProt IDQ00534
背景信息
功能： CDK6 (细胞周期蛋白依赖性激酶 6) 是一种丝氨酸/苏氨酸激酶，它与 D 型细胞周期蛋白形成活性复合物，并通过磷酸化视网膜母细胞瘤蛋白 (RB) 来调控 G1 期进程和 G1/S 期细胞周期转换，从而控制 E2F 依赖性转录程序 [1] [2] 。从机制上讲，CDK6 通过激活细胞周期蛋白 D 整合促有丝分裂信号和激素衍生信号，将细胞外生长信号与细胞周期启动和细胞增殖联系起来 [1] [3] 。除了经典的细胞周期调控外，越来越多的证据表明，CDK6 参与血液系统恶性肿瘤和实体瘤的发生发展，其中细胞周期蛋白 D-CDK6 信号通路失调会促进不受控制的增殖和治疗耐药性 [1] [4] 。在激素受体阳性乳腺癌的疾病模型中，异常的 CDK4/6 活性驱动 RB 磷酸化和细胞周期进程，使该通路成为重要的治疗靶点 [3] [5] 。与密切相关的亚型 CDK4 相比，CDK6 具有独特的调控和生物学功能，包括影响转录调控和肿瘤适应的激酶依赖性和非激酶依赖性活性 [4] [6] 。在实验应用中，选择性 CDK4/6 抑制剂，例如帕博西尼、瑞博西尼和阿贝西利，通过抑制 CDK4/6 活性来阻断 G1/S 期转换，尽管这些化合物对 CDK4 和 CDK6 的亲和力不同，并产生不同的生物学和临床效应 [3] [5] 。CDK6 过表达、含 CDK6 的耐药复合物以及 RB 相关通路的改变也与耐药机制有关，这支持了在癌症研究中继续探索 CDK6 选择性靶向策略 [5] [7] 。 亚细胞定位： 细胞质；细胞核；细胞突起，细胞皱褶；细胞质、细胞骨架、微管组织中心、中心体 表达水平： 组织特异性：广泛表达。在鳞状细胞癌、增殖性造血祖细胞、胰岛β细胞和神经母细胞瘤中积聚。在分化细胞中水平降低。 诱导：在肠道病毒 71 型 (EV71) 感染后表达下调。在 S 期进入期间由 NANOG 诱导。 亚基： 与 D 型 G1 期细胞周期蛋白相互作用。细胞周期蛋白结合通过 Thr-177 位点的磷酸化促进酶活化 (基于序列相似性)。与 RUNX1、CDKN2D、FBXO7 和 CDKN2C/p18-INK4c 结合。与 Hsp90/HSP90AB1 和 CDC37 形成胞质复合物。FBXO7 结合促进 D 型细胞周期蛋白的结合。与卡波西肉瘤疱疹病毒 (KSHV) V 细胞周期蛋白和猴疱疹病毒 (V 细胞周期蛋白/ECLF2) 相互作用；CDK6/V 细胞周期蛋白复合物磷酸化 NPM1，从而通过降低病毒 LANA 水平导致病毒再活化。