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| 货号 | PB9534 |
|---|---|
| 产品名称 | Anti-Cdk2 Antibody(原货号PB0562) |
| 基因名 | CDK2 |
| 抗体来源 | Rabbit |
| 克隆 | Polyclonal |
| 抗体亚型 | Rabbit IgG |
| 分子量 | 34KD |
| 免疫原 | E.coli-derived human Cdk2 recombinant protein (Position: E81-L298). Human Cdk2 shares 98.6% amino acid (aa) sequence identity with rat Cdk2. |
| 内容 | 500ug/ml;Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3. |
| 纯化方式 | Immunogen affinity purified. |
| 浓度 | 500ug/ml |
| 产品形态 | Lyophilized |
| 保存条件 | 12 months from date of receipt，-20℃ as supplied.6 months 2 to 8℃ after reconstitution.Avoid repeated freezing and thawing. |
| 背景资料 | CDK2, Cyclin-Dependent Kinase2, is also known as P33. The CDK2 protein was highly homologous to p34(CDC2) kinase and more significantly homologous to Xenopus Eg1 kinase, suggesting that CDK2 is the human homolog of Eg1. The CDK2 gene is mapped to 12q13, the same region to which the CDK4 gene maps. Human cyclin A binds independently to 2 kinases, p34(cdc2) or p33. In adenovirus-transformed cells, the viral E1A oncoprotein seems to associate with p33/cyclin A but not with p34(cdc2)/cyclin A. The gene for p33 shares 65% sequence identity with p34(cdc2). P33(cdk2) plays a unique role in cell cycle regulation of vertebrate cells. |
| 研究类别 | 1. Demetrick, D. J., Zhang, H., Beach, D. H.Chromosomal mapping of human CDK2, CDK4, and CDK5 cell cycle kinase genes.Cytogenet. Cell Genet. 66: 72-74, 1994.2. Tsai, L.-H., Harlow, E., Meyerson, M.Isolation of the human cdk2 gene that encodes the cyclin A- and adenovirus E1A-associated P33 kinase.Nature 353: 174-177, 1991. |
| Uniprot ID | CDK2: P24941 |
| 推荐配套的二抗和检测试剂 | Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P). *Blocking peptide 可以联系我们购买。 |
为了提供最优质的抗体，博士德对每一批抗体都用没有转染过的细胞系和体细胞组织检测，以保证博士德出品的抗体有足够的亲和性足以和对应蛋白天然的表达含量起反应。
| 应用 | 稀释度* |
|---|---|
| Western blot: | 1:500-2000 |
| Immunohistochemistry(Paraffin-embedded Section): | 1:50-400 |
| Immunocytochemistry/Immunofluorescence: | 1:50-400 |
*最优稀释度需要用户自己调试，此处数据仅供参考。
**博士德提供高敏感的二抗和检测试剂盒。详情见相关产品推荐。
Figure 1. Western blot analysis of Cdk2 using anti-Cdk2 antibody (PB9534). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: Rat Kidney Tissue LysateLane 2: Rat Liver Tissue LysateLane 3: Human Placenta Tissue LysateLane 4: HELA Whole Cell LysateLane 5: HUT Whole Cell LysateLane 6: JURKAT Whole Cell LysateLane 7: A549 Whole Cell Lysate After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Cdk2 antigen affinity purified polyclonal antibody (Catalog # PB9534) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Cdk2 at approximately 34KD. The expected band size for Cdk2 is at 34KD.
Figure 2. IHC analysis of Cdk2 using anti-Cdk2 antibody (PB9534).Cdk2 was detected in paraffin-embedded section of Mouse Testis Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Cdk2 Antibody (PB9534) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 3. IHC analysis of Cdk2 using anti-Cdk2 antibody (PB9534).Cdk2 was detected in paraffin-embedded section of Rat Testis Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Cdk2 Antibody (PB9534) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 4. IHC analysis of Cdk2 using anti-Cdk2 antibody (PB9534).Cdk2 was detected in paraffin-embedded section of Human Lung Cancer Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Cdk2 Antibody (PB9534) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 5. IF analysis of Cdk2 using anti- Cdk2 antibody (PB9534).
Cdk2 was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti- Cdk2 Antibody (PB9534) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 6. IF analysis of Cdk2 using anti- Cdk2 antibody (PB9534).
Cdk2 was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti- Cdk2 Antibody (PB9534) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
抗体的保存建议：抗体保存得当与否直接决定了抗体的活性和使用效果。如果抗体保存得当，大部分抗体活性都可以维持数月甚至数年。请按照说明书或者抗体标签上推荐的保存条件正确保存抗体！无论是保存还是运输，绝对避免反复冻融抗体！
1.拿到抗体后请务必在1000-3000转离心1-2分钟后再打开管盖进行分装和保存！（由于运输过程中反复颠簸，管内微量抗体容易聚集在管盖处，一旦直接打开容易流失抗体。请放心博士德抗体出厂时保证已装入足量。）
2.对于博士德大部分抗体，比较合适的保存方式是分装后保存在-20℃冰箱。
◇分装可以最大程度降低反复冻融对抗体活性的损害，同时也降低了多次从同一管中吸取抗体造成污染的可能性。
◇分装的量以一次实验用完为好，但建议最少不要少于10ul每份。分装体积越小，抗体浓度可能会受到蒸发以及管壁吸附的影响，同时分装次数越多移液吸头吸附的抗体也越多，可能会误认为抗体没有足量装入。（最好用无菌的进口0.2ML离心管来分装抗体，国产的小管一般没有经过很好的硅化处理，容易吸附抗体蛋白，影响抗体活性。）
◇分装时请将无菌的微量移液器吸头先插入管底部反复吹打几次后再吸出抗体，以便抗体有效成分与抗体保护剂充分混匀。
◇复融后的分装抗体如果一次用不完，请将剩余母液保存在4℃冰箱，避免再冻起来！
◇绝对避免将抗体保存在自动除霜冰箱的冷藏室中。尽量将抗体保存在手动除霜冰箱里面，而不是在冰箱门上。
3.大部分抗体收到后4℃短暂保存1-2周对抗体活性是基本没有影响的。如果抗体很快（1-2周内）就能使用完，推荐在4℃保存，避免反复冻融对抗体活性的损害。如果要长期保存则分装后-20℃保存。
4.即用型抗体请务必保存在4℃冰箱，一定不能冷冻保存。
5.特殊抗体的保存：
◇酶联抗体：一般保存在4℃，尽量避免冷冻起来。否则可能会导致酶活力的下降或者丧失。
◇偶联抗体：所有偶联抗体需要4℃避光保存。尤其是荧光标记抗体，对光极为敏感，因此所有实验阶段也是要避光操作的。
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