Revision 3

#13300

Store at -20C

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, eCLIP

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
105

Source/Isotype:
Rabbit IgG

UniProt ID:
#O75175

Entrez-Gene Id:
4849

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
eCLIP 1:200
For more information about the RBP-eCLIP service please visit Eclipsebio.

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

CNOT3 (E1L9S) Rabbit Monoclonal Antibody recognizes endogenous levels of total CNOT3 protein.

Species predicted to react based on 100% sequence homology

Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu638 of human CNOT3 protein.

Background

The evolutionarily conserved CCR4-NOT (CNOT) complex regulates mRNA metabolism in eukaryotic cells (1). This regulation occurs at different levels of mRNA synthesis and degradation, including transcription initiation, elongation, deadenylation, and degradation (1). Multiple components, including CNOT1, CNOT2, CNOT3, CNOT4, CNOT6, CNOT6L, CNOT7, CNOT8, CNOT9, and CNOT10 have been identified in this complex (2). In addition, subunit composition of this complex has been shown to vary among different tissues (3).

Research studies indicate that CNOT3 (along with CNOT1 and CNOT2) represses early developmental transcription factor expression, helping to maintain embryonic stem (ES) cell identity in mice and humans (4). Additional studies suggest that CNOT3 plays a role in mitotic progression as it destabilizes mitotic spindle assembly protein MAD1 mRNA (5). Finally, CNOT3 appears to act as a modifier gene affecting the penetrance of mutations causing autosomal dominant retinitis pigmentosa (6) and as a tumor suppressor associated with cases of adult T-cell acute lymphoblastic leukemia (7).

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

W: Western Blotting IP: Immunoprecipitation eCLIP: eCLIP

Cross-Reactivity Key

H: Human

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.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using CNOT3 (E1L9S) Rabbit mAb.
Western Blotting Image 1: CNOT3 (E1L9S) Rabbit Monoclonal Antibody
Western blot analysis of extracts from 293 cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human CNOT3 (hCNOT3-Myc/DDK; +), using CNOT3 (E1L9S) Rabbit mAb (upper), DYKDDDDK Tag (9A3) Mouse mAb #8146 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 2: CNOT3 (E1L9S) Rabbit Monoclonal Antibody
Immunoprecipitation of CNOT3 from PANC-1 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or CNOT3 (E1L9S) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using CNOT3 (E1L9S) Rabbit mAb.
Immunoprecipitation Image 1: CNOT3 (E1L9S) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Enhanced cross-linking and immunoprecipitation (eCLIP) was performed with RNA from K-562 cells and CNOT3 (E1L9S) Rabbit mAb using a protocol based on the RBP-eCLIP method from EclipseBio. The figure shows binding across the FBXO17 transcript. Data is kindly provided by the laboratory of Dr. Gene Yeo and used with permission.
eCLIP Image 1: CNOT3 (E1L9S) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.