Revision 3

#29786

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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, W-S, IP

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
23

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q9NWW6

Entrez-Gene Id:
54981

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
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

NRK1 (F4F8L) Rabbit Monoclonal Antibody recognizes endogenous levels of total NRK1 protein. This antibody does not cross-react with NRK2 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp48 of human NRK1 protein.

Background

Nicotinamide riboside (NR) is converted to NAD precursor nicotinamide mononucleotide (NMN) by nicotinamide riboside kinase 1 and 2 (NRK1 and NRK2), a function shared by yeast Nrk1 (1). Cytosolic NRK1, encoded by the NMRK1 gene, is expressed in most tissues and enriched in liver and kidney, while NRK2 expression is limited to muscle (2). Primary hepatocytes from NRK1 mice were used to demonstrate that NRK1 is essential for utilization of exogenous NR or NMN to produce NAD+ (2). Tissue-specific expression of NAD synthesis enzymes can drive cancer cell survival and treatment strategies. Cancer cells can arise in tissues with variable expression of rate-limiting enzymes for NAD synthesis, including nicotinate phosphoribosyltransferase (NAPRT) or nicotinamide phosphoribosyltransferase (NAMPT) (3). In instances where this expression is low, NRK1-driven NAD synthesis is key for cancer cell survival and may bypass inhibitors of the other NAD+ pathways (3).

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

W: Western Blotting W-S: Simple Western™ 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 3

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using NRK1 (F4F8L) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of NRK1 protein in A-673 and Kelly cells and negative expression in SH-SY5Y and SiMa cells is consistent with the predicted expression pattern.
Western Blotting Image 1: NRK1 (F4F8L) Rabbit Monoclonal Antibody
Western blot analysis of extracts from various tissues using NRK1 (F4F8L) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 2: NRK1 (F4F8L) Rabbit Monoclonal Antibody
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human NRK1 (hNRK1-Myc/DDK; +) or Myc/DDK-tagged full-length human NRK2 (hNRK2-Myc/DDK; +), using NRK1 (F4F8L) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 3: NRK1 (F4F8L) 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
Simple WesternTM analysis of lysates (0.1 mg/mL) from HDLM-2 cells using NRK1 (F4F8L) Rabbit mAb #29786. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western Blotting Image 1: NRK1 (F4F8L) Rabbit Monoclonal Antibody
Immunoprecipitation of NRK1 protein from HCT-15 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is NRK1 (F4F8L) Rabbit mAb. Western blot analysis was performed using NRK1 (F4F8L) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Immunoprecipitation Image 1: NRK1 (F4F8L) 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.