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PDP - Template Name: Monoclonal Antibody
PDP - Template ID: *******c5e4b77
R Recombinant
Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.

NADK Isoform 3 (F5E2T) Rabbit mAb #76851

Filter:
  • WB
Western Blotting Image 1: NADK Isoform 3 (F5E2T) Rabbit mAb
Western blot analysis of extracts from various cell lines using NADK Isoform 3 (F5E2T) Rabbit mAb (upper), NADK (E4R1P) Rabbit mAb #89833 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of NADK protein in MOLM-13, NCI-H292, and A-498 cells is consistent with the predicted expression pattern.

To Purchase # 76851

Cat. # Size Qty. Price Ships
76851S 100 µl
$370 Jan 22

Supporting Data

REACTIVITY H M R Mk
SENSITIVITY Endogenous
MW (kDa) 52
Source/Isotype Rabbit IgG
Application Key:
  • WB-Western Blotting 
Species Cross-Reactivity Key:
  • H-Human 
  • M-Mouse 
  • R-Rat 
  • Mk-Monkey 

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

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.

Protocol

Specificity / Sensitivity

NADK Isoform 3 (F5E2T) Rabbit mAb recognizes endogenous levels of total NADK isoform 3 protein (UniProt #O95544-3). This antibody does not cross-react with other NADK isoforms. This antibody cross-reacts with a 125 kDa protein of unknown identity in some cell lines.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ile49 of human NADK isoform 3 protein (Uniprot #O95544-3).

Background

NADK, also known as NAD kinase, is a cytoplasmic protein responsible for maintaining the pool of available NADP+ and NADPH within the cell (1). Using ATP as a phosphate donor, NADK catalyzes the phosphorylation of NAD+ to NADP+. This molecule is then reduced to NADPH and utilized in various metabolic and biosynthetic pathways (2). NADK has been suggested to play a role in glucose metabolism due to the effect NADPH production has on both the insulin secretion and survival of pancreatic β-cells (3). NADPH has a vital role in protecting cells from oxidative stress through its neutralizing effect on reactive oxygen species (ROS), which also accumulate during cell growth (2-4). Along with the p53 tumor suppression protein, NADK has been a suggested target in cancer therapy due to its link to NADPH production and its resulting protective role on growing and proliferating cells (2).

Cytoplasmic NADK isoform 3 is transcribed from a unique start site compared to isoforms 1 and 2, and lacks the N-terminal, Akt-dependent, activating phosphorylation sites of Ser44, Ser46, and Ser48 found in the other isoforms (5). Cells engineered to exclusively express NADK isoform 3 exhibit constitutively increased NAD kinase activity, insensitivity to AKT agonists, and increased synthesis of NADP+ (5). NADK isoform 3 is uniquely upregulated in metastatic nodules of a PDX model of melanoma (6). Isoform 3 expression is driven by transcription factor NRF2 in response to oxidative stress, allowing melanoma cells to survive both in vitro oxidative insult and in vivo metastatic cascade by increasing NADP+ synthesis (6).
For Research Use Only. Not For Use In Diagnostic Procedures.
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