Cat. # | Size | Qty. | Price | Inventory |
---|---|---|---|---|
33361T | 20 µl |
|
||
33361S | 100 µl |
|
REACTIVITY | H |
SENSITIVITY | Endogenous |
MW (kDa) | 28 |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunohistochemistry (Paraffin) | 1:100 - 1:400 |
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.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Do not allow slides to dry at any time during this procedure.
For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.
RECOMMENDED DETECTION REAGENTS |
SignalStain® Boost IHC Detection Reagent (HRP, Rabbit) #8114 | SignalStain® Boost IHC Detection Reagent (AP, Rabbit) #18653 |
---|---|---|
COMPATIBLE CHROMOGEN |
SignalStain® DAB Substrate Kit #8059 | SignalStain® Vibrant Red Alkaline Phosphatase Substrate Kit #76713 |
SignalStain® Vivid Purple Peroxidase Substrate Kit #96632 | SignalStain® Ultra Blue Alkaline Phosphatase Substrate Kit #12824 | |
SignalStain® Deep Black Peroxidase Substrate Kit #72986 | ||
SignalStain® Radiant Yellow Peroxidase Substrate Kit #69644 |
NOTE: Use of detection reagents other than those specified in this protocol may require further optimization of the primary antibody to account for the different sensitivities of the detection reagents.
posted February 2010
revised June 2020
Protocol Id: 283
Human
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro109 of human NNMT protein.
Nicotinamide N-methyltransferase (NNMT) is a metabolic enzyme expressed primarily in liver and adipose tissue. It catalyzes the transfer of a methyl group from S-Adenosyl-methionine (SAM) to nicotinamide, yielding 1-methylnicotinamide (MNAM) and S-Adenosyl-L-homocysteine (SAH) (1). This N-methylation enzymatic activity plays an important role in the biotransformation of drugs and xenobiotics, and also contributes to the metabolism of vitamin B3 (2). Knockdown of NNMT was shown to increase both SAM and NAD+ levels in white adipose tissue of high-fat diet-fed mice, resulting in increased energy expenditure and protection against diet-induced obesity (3). In contrast, increased liver NNMT expression in humans and mice correlated with an improved metabolic profile, through MNAM-mediated SIRT1 protein stabilization (4). In cancer cells, overexpression of NNMT resulted in excess consumption of methyl units from SAM, leading to histone hypomethylation that substantially altered the epigenetic landscape (5). These and other research studies have suggested that NNMT expression may have utility as a diagnostic and prognostic biomarker in cancer (6-8).
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