Revision 7

#67955Store 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-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

55

Source/Isotype:

Rabbit IgG

UniProt ID:

#P20248

Entrez-Gene Id:

890

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:800 - 1:3200
Immunofluorescence (Immunocytochemistry) 1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600

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 #29113.

Specificity / Sensitivity

Cyclin A2 (E6D1J) XP® Rabbit mAb recognizes endogenous levels of total cyclin A2 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human cyclin A2 protein. The epitope corresponds to a region surrounding Glu121 of human cyclin A2.

Background

While overcoming the G1/S checkpoint to commence DNA replication requires cyclin E, and traversing the G2/M checkpoint to initiate mitosis requires cyclin B to be present, cyclin A seems to be required for both S-phase and M-phase (1). A number of studies have described the ability of overexpressed cyclin A to accelerate the G1 to S transition, causing DNA replication, and cyclin A antisense DNA can prevent DNA replication (2-4). Cyclin A availability is apparently the rate-limiting step for entry into mitosis, and cyclin A is required for the completion of prophase (5). At late prophase, cyclin A may no longer be necessary as cdc2/cyclinB1 becomes active (5).

  1. Pagano, M. et al. (1992) EMBO. J. 11, 961-71.
  2. Resnitzky, D. et al. (1995) Mol. Cell. Biol. 15, 4347-52.
  3. d'Urso, G. et al. (1990) Science 250, 786-91.
  4. Zindy, F. et al. (1992) Biochem. Biophys. Res. Commun. 182, 1144-54.
  5. Furuno, N. et al. (1999) J. Cell. Biol. 147, 295-306.

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 BSA, 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-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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 7
#67955

Cyclin A2 (E6D1J) XP® Rabbit mAb

Western Blotting Image 1: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 Cyclin A2 (E6D1J) XP® Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对未经处理 (-) 或经过 Aphidicolin #32774(10 μg/mL,24 小时;+)或 Doxorubicin #5927(0.5 μM,24 小时;+)处理的各种人细胞系的提取物进行蛋白质印迹分析。正如预期的那样,Cyclin A2 蛋白被阿非迪霉素诱导并被阿霉素降低。Caki-1 细胞中Cyclin A2 蛋白的低表达与预测的表达模式一致。
Western Blotting Image 2: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 Cyclin A2 (E6D1J) XP® Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对转染带对照 siRNA (-) 或Cyclin A2 siRNA (+) 的 HCT 116 细胞提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
对 HCT 116 细胞提取物的细胞周期蛋白 A2 蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Cyclin A2 (E6D1J) XP® Rabbit mAb。使用 Cyclin A2 (E6D1J) XP® Rabbit mAb 进行蛋白印迹分析。小鼠Anti-兔 IgG (轻链特异) (D4W3E) 单克隆抗体 (HRP 偶联) #93702 用作二抗。
Immunohistochemistry Image 1: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 Cyclin A2 (E6D1J) XP® Rabbit mAb 对石蜡包埋的人尿路上皮癌进行免疫组织化学分析。
Immunohistochemistry Image 2: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 Cyclin A2 (E6D1J) XP® Rabbit mAb 对石蜡包埋的人结肠癌进行免疫组织化学分析。
Immunohistochemistry Image 3: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 Cyclin A2 (E6D1J) XP® Rabbit mAb 对石蜡包埋的人非小细胞肺癌进行免疫组织化学分析。
Immunohistochemistry Image 4: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 Cyclin A2 (E6D1J) XP® Rabbit mAb 对石蜡包埋的人非霍奇金氏淋巴瘤进行免疫组织化学分析。
Immunohistochemistry Image 5: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 Cyclin A2 (E6D1J) XP® Rabbit mAb 对石蜡包埋的人大细胞神经内分泌癌进行免疫组织化学分析。
Immunohistochemistry Image 6: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 Cyclin A2 (E6D1J) XP® Rabbit mAb 对石蜡包埋的人唾液腺小细胞癌组织进行免疫组织化学分析。
Immunohistochemistry Image 7: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 Cyclin A2 (E6D1J) XP® Rabbit mAb 对石蜡包埋的正常人胸腺进行免疫组织化学分析。
Immunohistochemistry Image 8: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)作为对照,使用 Cyclin A2 (E6D1J) XP® Rabbit mAb(左图)对石蜡包埋的人子宫内膜样腺癌进行免疫组织化学分析。
Immunohistochemistry Image 9: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 Cyclin A2 (E6D1J) XP® Rabbit mAb 对未经处理(左图)或已经过 Aphidicolin #32774(10 µg/ml,24 小时;右图)处理的石蜡包埋的 HT-29 细胞沉淀物进行免疫组织化学分析。
Immunofluorescence Image 1: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 Cyclin A2 (E6D1J) XP® Rabbit mAb(绿色)或 DyLight 650 Phalloidin #12956(红色)和 DAPI #4083(蓝色)对转染空载(左图,中等表达)、经过针对人细胞周期蛋白 A2 的 siRNA 转染(中图,低表达)或经过 Aphidicolin #32774(10 µg/mL,24 小时;右图,高表达)的 HCT 116 细胞进行共聚焦免疫荧光分析。
Flow Cytometry Image 1: Cyclin A2 (E6D1J) XP® Rabbit mAb Expand Image
使用 DRAQ5® #4084 和 Cyclin A2 (E6D1J) XP® Rabbit mAb(右图)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(左图)对 Jurkat 细胞进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488® Conjugate) #4412 用作二抗。