Revision 1

#25314

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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

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
130

Source/Isotype:
Rabbit IgG

UniProt ID:
#O75113

Entrez-Gene Id:
9683

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100

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

N4BP1 (E8H5E) Rabbit mAb recognizes endogenous levels of total N4BP1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu405 of human N4BP1 protein.

Background

N4BP1 (NEDD4-binding protein 1) was identified as a binding partner and substrate for the HECT domain-containing ubiquitin ligase NEDD4 (1). Subsequently, it was also found to interact with Itch, a ubiquitin ligase structurally similar to NEDD4 (2). Interaction between N4BP1 interferes with the ability of Itch to bind its substrates, including c-Jun and p63 (2). N4BP1 expression has been associated with immune regulation as well as cancer. N4BP1 was identified as an inhibitory factor of NF-κB-mediated MHC-1 expression in neuroblastoma and high expression of N4BP1 was linked to poor overall survival (3). Studies have also found that N4BP1 is a suppressor of cytokine production that can be cleaved and inactivated by caspase-8 (4). N4BP1 can interact with the NF-κB modulator NEMO to suppress NF-κB transcriptional activity (5). Studies have also identified N4BP1 as a potent interferon-inducible HIV-1 restriction factor that targets the degradation of viral mRNA (6).

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

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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human N4BP1 (hN4BP1-Myc/DDK; +), using N4BP1 (E8H5E) Rabbit Ab (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 1: N4BP1 (E8H5E) Rabbit mAb
Western blot analysis of extracts from various cells using N4BP1 (E8H5E) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of N4BP1 protein in MG-63 and OVSAHO cells is consistent with the predicted expression pattern.
Western Blotting Image 2: N4BP1 (E8H5E) Rabbit mAb
Immunoprecipitation of N4BP1 protein from COV434 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is N4BP1 (E8H5E) Rabbit mAb. Western blot analysis was performed using N4BP1 (E8H5E) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation Image 1: N4BP1 (E8H5E) Rabbit mAb
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.