Revision 1

#74736Store 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

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

45

SOURCE:

Rabbit

UniProt ID:

#P11157

Entrez-Gene Id:

20135

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50

Storage

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

Specificity / Sensitivity

Phospho-RRM2 (Thr33) Antibody recognizes endogenous levels of RRM2 protein only when phosphorylated at Thr33. By western blot, this antibody detects a 110 kDa band of unknown origin.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr33 of mouse RRM2 protein. Antibodies are purified by peptide affinity chromatography.

Background

Ribonucleotide reductase catalyzes the rate-limiting step in the synthesis of deoxynucleotide triphosphates (dNTPs). Ribonucleoside-diphosphate reductase subunit M2 (RRM2) is frequently overexpressed and associated with poor prognosis in multiple human cancers (1). RRM2/AKT/NF-κB signaling pathway is implicated in tumor invasiveness in gastric cancer (2). RRM2 is highly expressed in melanoma, and correlated with poor prognosis in BRAF-mutant melanoma. Knockdown of RRM2 stabilized the transient response of cells and patient-derived xenograft (PDX) model system to BRAF inhibition (3).
Cyclin-dependent kinase (CDK) mediated phosphorylation of RRM2 at Thr33 targets the protein for degradation, allowing cells to maintain balanced dNTP pools (4).

  1. Furuta, E. et al. (2010) Biochim Biophys Acta 1805, 141-52.
  2. Zhong, Z. et al. (2016) Pharmazie 71, 280-4.
  3. Fatkhutdinov, N. et al. (2016) Mol Cancer Res 14, 767-75.
  4. D'Angiolella, V. et al. (2012) Cell 149, 1023-34.

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

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.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#74736

Phospho-RRM2 (Thr33) Antibody

Western Blotting Image 1: Phospho-RRM2 (Thr33) Antibody Expand Image
Western blot analysis of extracts from PANC-1 and C2C12 cells, untreated (-) or treated with Roscovitine #9885 (20 μM, 8 hr; +), using Phospho-RRM2 (Thr33) Antibody (upper), RRM2 (E7Y9J) XP® Rabbit mAb #65939 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Phosphorylation of RRM2 protein at Thr33 is reduced by roscovitine treatment as expected.
Western Blotting Image 2: Phospho-RRM2 (Thr33) Antibody Expand Image
Western blot analysis of extracts from C2C12 and KNRK cells, untreated (-) or treated with λ phosphatase/calf intestinal phosphatase (CIP) (+), using Phospho-RRM2 (Thr33) Antibody (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Immunoprecipitation Image 1: Phospho-RRM2 (Thr33) Antibody Expand Image
Immunoprecipitation of phospho-RRM2 (Thr33) protein from C2C12 cell extracts. Lane 1 is 10% input, lane 2 is Normal Rabbit IgG #2729, and lane 3 is Phospho-RRM2 (Thr33) Antibody. Western blot analysis was performed using Phospho-RRM2 (Thr33) Antibody. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.