Anti-Tissue Transglutaminase Antibody, FN binding domain, clone 4G3

Tissue transglutaminase (tTG) is a 75 kDa monomeric globular protein expressed in the majority of cells and tissues. tTG localizes mainly in the cytoplasm but some tGT is present on the cell surface and is association with the extracellular matrix. In addition to its guanosine triphosphatase (GTPase) activity, tTG catalyzes the posttranslational modification of proteins by transamidation of available glutamine residues (Monsonego, 1997). Extracellular tTG is able to bind to and cross-link several ECM proteins and may function to stabilize the ECM and basement membranes. The protein is translated as a fully active enzyme and there is no evidence for proteolytic activation.

High constitutive expression and accumulation of active tTG is observed in (among others) endothelial cells, platelets, vascular smooth muscle cells and epithelial cells of the lens. tTG is expressed at very low levels in skeletal muscle cells and neurons and may be difficult to detect in these cell/tissue types.

Tissue Transglutaminase. Will recognize all three known splice variants of tTG.

Epitope: FN binding domain

Fibronectin binding domain of tissue transglutaminase (amino acids 1-165 of tTG).

Detect Tissue Transglutaminase using this Anti-Tissue Transglutaminase Antibody, FN binding domain, clone 4G3 validated for use in IC.

Immunohistochemistry: 20 μg/mL

Adhesion Blocking: 10-20 μg/mL (per 2x10e5 cells). 4G3 interferes with adhesion of TPA stimulated THP-1 cells and M-CSF stimulated peripheral blood monocytes to fibronectin.

Optimal working dilutions must be determined by the end user.



PROTOCOL:

IMMUNOCYTOCHEMISTRY: THP-1 cells were induced to differentiate into macrophages by treatment with 150 ng/ml of TPA and plated on fibronectin coated glass coverslips for 72 hours.Live non-permeablized cells were incubated with 20 μg/mL of MAB3839 for 45 minutes, washed three times with PBS and fixed for 10 minutes with 3.7% paraformaldehyde. The cells were then washed with PBS and incubated 45 minutes with a fluoresceine labeled affinity-purified goat-anti-mouse secondary antibody. After extensive washing with PBS, cells were mounted in Fluorsave medium.

Observed Staining:Cell surface tTG was colocalized with b1- and b3-Integrins in podosome-like adhesive structures of macrophages adherent on fibronectin.

Research Category
Apoptosis & Cancer

Metabolism

Research Sub Category
Apoptosis - Additional

Enzymes & Biochemistry

Format: Purified

Protein A purified immunoglobulin in 0.02M Phosphate Buffer, pH 7.6, 0.25M NaCl containing 0.1% sodium azide.

Maintain at 2-8°C in undiluted aliquots for up to 12 months.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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