Revision 5

#18048Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-F, IF-IC

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

33

Source/Isotype:

Rabbit IgG

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:200 - 1:800
Immunofluorescence (Frozen) 1:400 - 1:1600
Immunofluorescence (Immunocytochemistry) 1:800 - 1:3200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #34541.

Specificity / Sensitivity

CDK2 (E8J9T) XP® Rabbit mAb recognizes endogenous levels of total CDK2 protein. In rodent cell lines, isoforms 1 and 2 of CDK2 run as two distinct bands by western blot. This antibody non-specifically labels the muscle in the mouse retina and the vasculature in the mouse brain by immunofluorescence.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro294 of human CDK2 protein.

Background

Cyclin-dependent kinase 2 (p33CDK2) is an important component of the cell cycle machinery. Like p34cdc2, kinase activity is regulated by phosphorylation state as well as association with a cyclin subunit and a CDK inhibitor. Inhibitory phosphorylation occurs on Thr14 and Tyr15 (1). Inhibition of CDK2-cyclin complexes can also be attributed to association with p27 Kip1 and p21 Waf1/Cip1 (2). Activation of CDK2 complexes requires dephosphorylation of Thr14 and Tyr15 by cdc25 phosphatase and phosphorylation of Thr160 (3), which is mediated by CAK, a complex of CDK7 and cyclin H (4). CDK2/cyclin E kinase activity is important for the G1 to S transition and phosphorylation of the Rb protein. During S-phase, active CDK2/cyclin A complexes predominate and phosphorylate E2F and the active CDK2 complex persists in the nucleus throughout G2 (5).

  1. Morgan, D.O. (1995) Nature 374, 131-4.
  2. Poon, R.Y. et al. (1996) J Biol Chem 271, 13283-91.
  3. Gu, Y. et al. (1992) EMBO J 11, 3995-4005.
  4. Fesquet, D. et al. (1993) EMBO J 12, 3111-21.
  5. Morgan, D.O. (1997) Annu Rev Cell Dev Biol 13, 261-91.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalStain is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 5
#18048

CDK2 (E8J9T) XP® Rabbit mAb

Western Blotting Image 1: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
使用 CDK2 (E8J9T) XP®(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对不同细胞系提取物进行蛋白质印迹法分析。
Immunoprecipitation Image 1: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
对 SK-MEL-30 细胞提取物 CDK2 蛋白进行免疫沉淀。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 CDK2 (E8J9T) XP® Rabbit mAb。使用 CDK2 (E8J9T) XP® Rabbit mAb 进行蛋白质印迹法分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Immunohistochemistry Image 1: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
使用 CDK2 (E8J9T) XP® Rabbit mAb 对石蜡包埋的正常人结肠进行免疫组织化学分析。
Immunohistochemistry Image 2: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
使用 CDK2 (E8J9T) XP® Rabbit mAb (top) or CDK2 Rabbit mAb(底部)对石蜡包埋的人卵巢癌(左图)或结肠癌(右图)细胞进行免疫组织化学分析。这两种抗体可检测人 CDK2 上的独立、独特表位。使用两种抗体获得的相似染色模式有助于确认染色特异性。
Immunohistochemistry Image 3: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
以浓度匹配的兔 (DA1E) 单克隆抗体 IgG XP® 同型对照 #3900(右图)作为对照,使用 CDK2 (E8J9T) XP® Rabbit mAb(左图)对石蜡包埋的人食管腺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 4: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
使用 CDK2 (E8J9T) XP® Rabbit mAb 对石蜡包埋的 SK-MEL-30 细胞沉淀物(左图,高表达)或 Hep G2 细胞沉淀物(右图,低表达)进行免疫组织化学分析。
Immunohistochemistry Image 5: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
使用 CDK2 (E8J9T) XP® Rabbit mAb 对石蜡包埋的正常小鼠胃进行免疫组织化学分析。
Immunohistochemistry Image 6: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
使用 CDK2 (E8J9T) XP® Rabbit mAb(顶图)或 CDK2 Rabbit mAb(底图),对石蜡包埋的正常小鼠睾丸(左)或子宫(右)进行免疫组织化学分析。这两种抗体可检测人 CDK2 上的独立、独特表位。使用两种抗体获得的相似染色模式有助于确认染色特异性。
Immunohistochemistry Image 7: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
使用 CDK2 (E8J9T) XP® Rabbit mAb 对石蜡包埋的野生型 HCT116 细胞沉淀物(左图,阳性)或敲除型 CDK2 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Immunofluorescence Image 1: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
使用 CDK2 (E8J9T) XP® Rabbit mAb(绿色)对小鼠结肠进行共聚焦免疫荧光分析。在用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭游离次要结合位点后,用 Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182(红色色)对组织进行共同标记。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Immunofluorescence Image 2: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
使用 CDK2 (E8J9T) XP® Rabbit mAb(绿色)对小鼠胸腺进行共聚焦免疫荧光分析。在用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭游离次要结合位点后,用 Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182(红色色)对组织进行共同标记。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Immunofluorescence Image 1: CDK2 (E8J9T) XP® Rabbit mAb Expand Image
使用 CDK2 (E8J9T) XP® Rabbit mAb(绿色)、DyLight 554 Phalloidin #13054(红色)和 DAPI #4083(蓝色)对 HCT 116 细胞(左图,阳性)或 CDK2 HCT 116 敲除型细胞(右图,阴性)进行共聚焦免疫荧光分析。