REACTIVITY | H M R |
SENSITIVITY | Endogenous |
MW (kDa) | 4-12 |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunohistochemistry (Paraffin) | 1:75 |
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, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with recombinant protein corresponding to the full-length human CART protein.
Cocaine- and amphetamine-regulated transcript (CART) peptides are neurotransmitters of 39 and 47 amino acids that are involved in a variety of physiological processes. The CART precursor, a polypeptide of 116 residues, requires prohormone/proprotein convertase-mediated endoproteolytic cleavage to produce the two active peptides (1). CART peptides are found in several neuroendocrine tissues such as the brain, pituitary, adrenals, and pancreas (2). Hypothalamic CART is regulated by leptin, and plays a role in appetite and feeding behavior (3). Mesolimbic CART is regulated by CREB and may play a role in drug abuse behaviors by mediating some of CREB effects (4). Pancreatic CART is found in islet endocrine cells and parasympathetic and sensory nerves. It inhibits glucose-stimulated insulin secretion and has been found to be up-regulated in beta cells in animal model of diabetes (5). A missense mutation in the corresponding CART gene can correlate with susceptibility to obesity and reduced resting energy expenditure (6).
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