Revision 3

#97293Store 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, W-S, IP

REACTIVITY:

M

SENSITIVITY:

Endogenous

MW (kDa):

45

Source/Isotype:

Rabbit IgG

UniProt ID:

#P11157

Entrez-Gene Id:

20135

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50

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.

Specificity / Sensitivity

RRM2 (F4O1C) Rabbit mAb recognizes endogenous levels of total RRM2 protein.

Species Reactivity:

Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of mouse RRM2 protein.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ 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 3
#97293

RRM2 (F4O1C) Rabbit mAb

Western Blotting Image 1: RRM2 (F4O1C) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length mouse RRM2 (mRRM2-Myc/DDK; +), using RRM2 (F4O1C) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 2: RRM2 (F4O1C) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using RRM2 (F4O1C) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of RRM2 protein in Neuro-2a cells is consistent with the predicted expression pattern.
Western Blotting Image 1: RRM2 (F4O1C) Rabbit mAb Expand Image
Simple Western™ analysis of lysates (0.1 mg/mL) from C2C12 cells using RRM2 (F4O1C) Rabbit mAb #97293. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Immunoprecipitation Image 1: RRM2 (F4O1C) Rabbit mAb Expand Image
Immunoprecipitation of RRM2 protein from C2C12 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is RRM2 (F4O1C) Rabbit mAb. Western blot analysis was performed using RRM2 (F4O1C) Rabbit mAb. Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 was used as a secondary antibody.