Revision 5

#55506Store 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-Bond, IHC-P, IF-IC, FC-FP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

36

Source/Isotype:

Rabbit IgG

UniProt ID:

#P24385

Entrez-Gene Id:

595

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:250 - 1:1000
Immunohistochemistry (Paraffin) 1:250 - 1:1000
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 #66467.

Specificity / Sensitivity

Cyclin D1 (E3P5S) XP® Rabbit mAb recognizes endogenous levels of total cyclin D1 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala284 of human cyclin D1 protein.

Background

Activity of the cyclin-dependent kinases CDK4 and CDK6 is regulated by T-loop phosphorylation, by the abundance of their cyclin partners (the D-type cyclins), and by association with CDK inhibitors of the Cip/Kip or INK family of proteins (1). The inactive ternary complex of cyclin D/CDK4 and p27 Kip1 requires extracellular mitogenic stimuli for the release and degradation of p27 concomitant with a rise in cyclin D levels to affect progression through the restriction point and Rb-dependent entry into S-phase (2). The active complex of cyclin D/CDK4 targets the retinoblastoma protein for phosphorylation, allowing the release of E2F transcription factors that activate G1/S-phase gene expression (3). Levels of cyclin D protein drop upon withdrawal of growth factors through downregulation of protein expression and phosphorylation-dependent degradation (4).

  1. Hirai, H. et al. (1995) Mol Cell Biol 15, 2672-81.
  2. Sherr, C.J. (1996) Science 274, 1672-7.
  3. Lukas, J. et al. (1996) Mol Cell Biol 16, 6917-25.
  4. Diehl, J.A. et al. (1997) Genes Dev 11, 957-72.

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-Bond: IHC Leica Bond 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 5
#55506

Cyclin D1 (E3P5S) XP® Rabbit mAb

Western Blotting Image 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
使用 Cyclin D1 (E3P5S) XP® Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对不同细胞系的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
用 Cyclin D1 对 SH-SY5Y 细胞提取物进行免疫沉淀。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Cyclin D1 (E3P5S) XP® Rabbit mAb。使用作为一抗的 Cyclin D1 (E3P5S) XP® Rabbit mAb 和作为二抗的 Anti-rabbit IgG, HRP-linked Antibody #7074 进行蛋白质印迹法分析。
Immunohistochemistry Image 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 Cyclin D1 (E3P5S) XP® Rabbit mAb 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 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
使用 Cyclin D1 (E3P5S) XP® Rabbit mAb 对石蜡包埋的人尿路上皮癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
使用 Cyclin D1 (E3P5S) XP® Rabbit mAb(左图)或 Cyclin D1 (92G2) Rabbit mAb #2978(右图)对石蜡包埋的人乳腺导管癌细胞进行免疫组织化学分析。这两种抗体可检测人Cyclin D1 上的独立、特异的表位。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: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
使用 Cyclin D1 (E3P5S) XP® Rabbit mAb 对石蜡包埋的人肺腺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 4: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
使用 Cyclin D1 (E3P5S) XP® Rabbit mAb 对石蜡包埋的小鼠肺 4T1 转移瘤细胞进行免疫组织化学分析。
Immunohistochemistry Image 5: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
使用 Cyclin D1 (E3P5S) XP® Rabbit mAb 对石蜡包埋的人卵巢癌肉瘤细胞进行免疫组织化学分析。
Immunohistochemistry Image 6: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® 同型对照 #3900(右图)作为对照,使用 Cyclin D1 (E3P5S) XP® Rabbit mAb(左图)对石蜡包埋的小鼠胰腺细胞进行免疫组织化学分析。
Immunohistochemistry Image 7: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
使用 Cyclin D1 (E3P5S) XP® Rabbit mAb 对石蜡包埋的 SH-SY5Y 细胞(左图,阳性)或 THP-1 细胞团块(右图,阴性)进行免疫组织化学分析。
Immunofluorescence Image 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
使用 Cyclin D1 (E3P5S) XP® Rabbit mAb(绿色)对 SH-SY5Y 细胞(左图,阳性)和 293T 细胞(右图,阴性)进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。
Flow Cytometry Image 1: Cyclin D1 (E3P5S) XP® Rabbit mAb Expand Image
使用 Cyclin D1 (E3P5S) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® 同型对照 #3900(虚线)对 K-562 细胞(蓝色)和 SH-SY5Y 细胞(绿色)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。