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.
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UniProt ID:

#Q96GD4

Entrez-Gene Id:

9212

Product Information

Storage

Antibodies are supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Do not aliquot the antibodies. Peptides are supplied at 6 µM in 0.001% DMSO.
Enzyme is supplied in 50 mM Tris-HCl, pH7.5; 150 mM NaCl, 0.25 mM DTT, 0.1 mM EGTA, 0.1 mM EDTA, 0.1 mM PMSF, 25% glycerol, 7 mM glutathione.
Store at –80°C.
Keep enzymes on ice during use.
Avoid repeated freeze-thaw cycles.

Source / Purification

The GST-Kinase fusion protein was produced using a baculovirus expression system with a construct expressing full-length human Aurora B (Met1-Ala344) (GenBank Accession No. NM_004217) with an amino-terminal GST tag. The protein was purified by one-step affinity chromatography using GSH-agarose.

Product Description

The kit provides a means of performing kinase activity assays with recombinant human Aurora B kinase. It includes active Aurora B kinase (supplied as a GST fusion protein), a biotinylated peptide substrate and a phospho-serine antibody for detection of the phosphorylated form of the substrate peptide.
Molecular Formula Biotin-PLK (Ser137): 1,945 Daltons. GST-Aurora B Kinase: 66 kDa.

Background

Aurora kinases belong to a highly conserved family of mitotic serine/threonine kinases with three members identified among mammals: Aurora A, B, and C (1,2). Studies on the temporal expression pattern and subcellular localization of Aurora kinases in mitotic cells suggest an association with mitotic structure. Aurora kinase functional influences span from G2 phase to cytokinesis and may be involved in key cell cycle events such as centrosome duplication, chromosome bi-orientation and segregation, cleavage furrow positioning, and ingression (3). Aurora A is detected at the centrosomes, along mitotic spindle microtubules, and in the cytoplasm of mitotically proliferating cells. Aurora A protein levels are low during G1 and S phases and peak during the G2/M phase of the cell cycle. Phosphorylation of Aurora A at Thr288 in its catalytic domain increases kinase activity. Aurora A is involved in centrosome separation, maturation, and spindle assembly and stability. Expression of Aurora B protein also peaks during the G2/M phase of the cell cycle; Aurora B kinase activity peaks at the transition from metaphase to the end of mitosis. Aurora B associates with chromosomes during prophase prior to relocalizing to the spindle at anaphase. Aurora B regulates chromosome segregation through the control of microtubule-kinetochore attachment and cytokinesis. Expression of both Aurora A and Aurora B during the G2/M phase transition is tightly coordinated with histone H3 phosphorylation (4,5); research investigators have observed overexpression of these kinases in a variety of human cancers (2,4). Aurora C localizes to the centrosome from anaphase to cytokinesis and both mRNA and protein levels peak during G2/M phase. Although typical Aurora C expression is limited to the testis, research studies report overexpression of Aurora C is detected in various cancer cell lines (6).

  1. Warner, S.L. et al. (2003) Mol Cancer Ther 2, 589-95.
  2. Katayama, H. et al. (2003) Cancer Metastasis Rev 22, 451-64.
  3. Andrews, P.D. et al. (2003) Curr Opin Cell Biol 15, 672-83.
  4. Pascreau, G. et al. (2003) Prog Cell Cycle Res 5, 369-74.
  5. Crosio, C. et al. (2002) Mol Cell Biol 22, 874-85.
  6. Kimura, M. et al. (1999) J Biol Chem 274, 7334-40.

Species Reactivity

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

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.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

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Revision 1
#7513

HTScan® Aurora B Kinase Assay Kit

HTScan® Aurora B Kinase Assay Kit: Image 1 Expand Image
图 3. Dose dependence curve of Aurora B kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of substrate peptide #1300 by Aurora B kinase. 在 50 µl 反应体积中,每个反应孔在室温下用数量递增的 Aurora B 和 1.5 µM 底物肽反应 30 分钟。(DELFIA® 是 PerkinElmer, Inc. 的注册商标)
HTScan® Aurora B Kinase Assay Kit: Image 2 Expand Image
图 5. Staurosporine inhibition of Aurora B kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of Aurora B substrate peptide #1300 by Aurora B kinase. 在 50 µl 反应体积中,每个反应孔在室温下用 50 ng Aurora B、1.5 µM 底物肽、50 µM ATP 和数量递增的 staurosporine 反应 30 分钟。(DELFIA® 是 PerkinElmer, Inc. 的注册商标)
HTScan® Aurora B Kinase Assay Kit: Image 3 Expand Image
图 4. Peptide concentration dependence of Aurora B kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of substrate peptide #1300 by Aurora B kinase. 在 50 µl 反应体积中,每个反应孔在室温下用 50 ng Aurora B 和浓度递增的底物肽反应 30 分钟。(DELFIA® 是 PerkinElmer, Inc. 的注册商标)
HTScan® Aurora B Kinase Assay Kit: Image 4 Expand Image
图 2. Time course of Aurora B kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of Aurora B substrate peptide #1300 by Aurora B kinase. 在 50 µl 反应体积中,每个反应孔用 50 ng Aurora B 和 1.5 µM 底物肽。(DELFIA® 是 PerkinElmer, Inc. 的注册商标)
HTScan® Aurora B Kinase Assay Kit: Image 5 Expand Image
图 1. 使用以下反应条件在放射分析中测量 Aurora B 激酶活性:5 mM MOPS、pH 7.2、2.5 mM β-甘油磷酸酯、1 mM EGTA、0.4 mM EDTA、5 mM MgCl2、0.05 mM DTT、50 μM ATP,底物:MBP 200 ng/μL,和重组 Aurora B:变量。