Revision 3

#63409Store 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, eCLIP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

100

Source/Isotype:

Rabbit IgG

UniProt ID:

#O15226

Entrez-Gene Id:

55922

Product Information

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

NKRF (E9G1T) Rabbit mAb recognizes endogenous levels of total NKRF protein.

Species Reactivity:

Human, Mouse, Rat

Species predicted to react based on 100% sequence homology

Dog, Pig

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro75 of human NKRF protein.

Background

NF-κB Repressing Factor (NKRF) is a constitutively expressed transcription factor that binds to the negative regulatory element within the promotor region of many genes that are transactivated by NF-κB (1). Indeed, research studies have found NKRF to repress transcription of IL-8/CXCL8, iNOS, CXCL10, and IFN-γ (2-4). In addition to regulating the expression of pro-inflammatory factors, NKRF has been shown to play a role in cell survival in the context of proteotoxic stress, through the regulation of rRNA processing and ribosome biogenesis (5).

  1. Nourbakhsh, M. and Hauser, H. (1999) EMBO J 18, 6415-25.
  2. Nourbakhsh, M. et al. (2001) J Biol Chem 276, 4501-8.
  3. Feng, X. et al. (2002) Proc Natl Acad Sci U S A 99, 14212-7.
  4. Huang, K.H. et al. (2013) PLoS One 8, e77789.
  5. Coccia, M. et al. (2017) Proc Natl Acad Sci U S A 114, 1045-1050.

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 IP: Immunoprecipitation eCLIP: eCLIP

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.
XP is a registered 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
#63409

NKRF (E9G1T) Rabbit mAb

Western Blotting Image 1: NKRF (E9G1T) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using NKRF (E9G1T) Rabbit mAb.
Western Blotting Image 2: NKRF (E9G1T) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, transfected with control siRNA (-) or NKRF siRNA (+), using NKRF (E9G1T) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). The loss of signal in extracts from NKRF siRNA-transfected cells confirms specificity of the antibody for NKRF.
Immunoprecipitation Image 1: NKRF (E9G1T) Rabbit mAb Expand Image
Immunoprecipitation of NKRF protein from A549 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is NKRF (E9G1T) Rabbit mAb. Western blot analysis was performed using NKRF (E9G1T) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
eCLIP Image 1: NKRF (E9G1T) Rabbit mAb Expand Image
Enhanced cross-linking and immunoprecipitation (eCLIP) was performed with RNA from K-562 cells and NKRF (E9G1T) Rabbit mAb using a protocol based on the RBP-eCLIP method from Eclipsebio. The figure shows binding across the NKRF transcript. Data is kindly provided by the laboratory of Dr. Gene Yeo and used with permission.