Revision 1
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, IHC-P, IF-IC

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

48

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q13309

Entrez-Gene Id:

6502

Product Information

Product Usage Information

This product is the carrier free version of product #2652. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

Formulation

Supplied in 1X PBS, BSA and Azide Free.

For standard formulation of this product see product #2652

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity / Sensitivity

Skp2 (D3G5) XP® Rabbit mAb (BSA and Azide Free) recognizes endogenous levels of total Skp2 protein. This antibody is predicted to cross-react with Skp2α and Skp2β.

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human Skp2 protein.

Background

Members of the F-box family of proteins are characterized by the approximate 40 amino acid F-box motif named after cyclin F (1,2). F-box proteins constitute one of the four subunits of the Skp1-Cullin-F-box (SCF) ubiquitin ligase complex. The substrate specificity of SCF complexes is determined by the interchangeable F-box proteins, which act as adaptors by associating with phosphorylated substrate proteins and recruiting them to the SCF core. F-box proteins contain two fundamental domains: the F-box motif mediates binding to Skp1 and a leucine rich repeat (LRR) domain mediates substrate interactions.
S phase kinase-associated protein 2 (Skp2) interacts with cyclin A/CDK2 and mediates G1 to S and G2 to M phase transitions by targeting the cyclin-dependent kinase (CDK) inhibitors p27, p21, and p130 for ubiquitination and subsequent proteolysis (3-6). Overexpression of Skp2 results in deregulated proliferation and genetic instabilities typical of cancer cells (7). Research studies have shown that increased Skp2/decreased p27 levels are associated with many aggressive lymphomas and human carcinomas such as colon, breast, prostate and lung cancers (7). Several recent research studies have demonstrated that Skp2 is subject to phosphorylation-dependent regulation by a network of pro-proliferative Ser/Thr kinases. It appears as though phosphorylation of Skp2 at Ser64 by CDK2 (8), Ser72 by Akt1 (9), and Thr417 by PIM1 (10) promotes stabilization of Skp2, possibly constituting an additional mechanism for Skp2 oncogenicity.

  1. Pagano, M. (2004) Mol Cell 14, 414-6.
  2. Reed, S.I. (2003) Nat Rev Mol Cell Biol 4, 855-64.
  3. Zhang, H. et al. (1995) Cell 82, 915-25.
  4. Nakayama, K. et al. (2004) Dev Cell 6, 661-72.
  5. Bornstein, G. et al. (2003) J Biol Chem 278, 25752-7.
  6. Tedesco, D. et al. (2002) Genes Dev 16, 2946-57.
  7. Bloom, J. and Pagano, M. (2003) Semin Cancer Biol 13, 41-7.
  8. Rodier, G. et al. (2008) EMBO J 27, 679-91.
  9. Gao, D. et al. (2009) Nat Cell Biol 11, 397-408.
  10. Cen, B. et al. (2010) J Biol Chem 285, 29128-37.

Species Reactivity

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

Applications Key

WB: Western Blotting IHC-P: Immunohistochemistry (Paraffin) 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 1
#95000

Skp2 (D3G5) XP® Rabbit mAb (BSA and Azide Free)

Western Blotting Image 1: Skp2 (D3G5) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Skp2 (D3G5) XP® Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。数据的生成使用的是该产品的常规组分产品。
Western Blotting Image 2: Skp2 (D3G5) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Skp2 (D3G5) XP® Rabbit mAb, 对转染空载 (-) 或转染能编码全长人 Skp2 的带有 Myc/DDK 标签的 cDNA 表达载体 (hSkp2-Myc/DDK, +) 的 293T 细胞提取物进行蛋白质印迹分析。数据的生成使用的是该产品的常规组分产品。
Western Blotting Image 3: Skp2 (D3G5) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Skp2 (D3G5) XP® Rabbit mAb(上图)和 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对表达非靶向 shRNA (shNT) 或 shSkp2 的 HeLa 细胞提取物进行蛋白质印迹分析。数据的生成使用的是该产品的常规组分产品。
Immunohistochemistry Image 1: Skp2 (D3G5) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Skp2 (D3G5) XP® Rabbit mAb 对石蜡包埋的人肺癌细胞进行免疫组织化学分析。数据的生成使用的是该产品的常规组分产品。
Immunohistochemistry Image 2: Skp2 (D3G5) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Skp2 (D3G5) XP® Rabbit mAb 对石蜡包埋的人淋巴瘤细胞进行免疫组织化学分析。数据的生成使用的是该产品的常规组分产品。
Immunohistochemistry Image 3: Skp2 (D3G5) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Skp2 (D3G5) XP® Rabbit mAb 对石蜡包埋的人卵巢癌细胞进行免疫组织化学分析。数据的生成使用的是该产品的常规组分产品。
Immunofluorescence Image 1: Skp2 (D3G5) XP® Rabbit mAb (BSA and Azide Free) Expand Image
使用 Skp2 (D3G5) XP® Rabbit mAb(绿色),对表达非靶向 shRNA(shNT;左图)或 Skp2(右图)的 HeLa 细胞进行共聚焦免疫荧光分析。肌动蛋白丝用 DY-554 phalloidin(红色)标记(细胞系由哈佛大学医学院的 Wenyi Wei 博士惠赠)。数据的生成使用的是该产品的常规组分产品。