Cat. # | Size | Qty. | Price |
---|---|---|---|
3557T | 20 µl |
|
|
3557S | 100 µl |
|
REACTIVITY | H M R Mk |
SENSITIVITY | Endogenous |
MW (kDa) | 78 |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunohistochemistry (Paraffin) | 1:100 |
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, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys598 of human TGM2.
Transglutaminase 2 (TGM2) is a calcium-dependent enzyme that cross-links both cytosolic and extracellular matrix proteins by catalyzing the formation of bonds between lysine and glutamine residues (1). This bifunctional enzyme also has intrinsic GTPase activity, and it has been suggested that regulation of the transamidase activity might be regulated through a G-protein coupled receptor-signaling pathway (2). In cross-linking peptides, TGM2 helps to regulate cytoskeletal structure, cell migration, apoptosis and cell-matrix adhesion. In addition, the enzyme plays an important role in wound healing and the immune response (3). TGM2 has exhibited kinase activity in vitro, with insulin-like growth factor-binding protein-3 (IGFBP-3) as one possible substrate (4). This widely expressed protein is localized to the cytosol and nucleus, but has also been isolated from the cell surface and extracellular matrix (reviewed in 5). Because of its interaction with a number of different substrates, and its role in the response to injury, TGM2 has been associated with the pathology of a number of human disorders. It has long been recognized as the major autoantigen in celiac disease (6); altered TGM2 expression or activity may be associated with Alzheimer disease, Huntington disease, arteriosclerosis, diabetes, and numerous forms of cancer (reviewed in 7).
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