Transglutaminase 2

Art. No.

A014

Synonym

Polyclonal antibody to human tissue transglutaminase (TG2)

Background info

Tissue transglutaminase is a, Ca2+-dependent enzyme (78 kDa) composed by 4 domains: Beta Sheet Domain (fibronectin binding, ~17 kDa), catalytic Core Domain (Cys-His-Asp catalytic triad, Calcium-binding, GTP/GDP-binding, ~37 kDa), Beta Barrel 1 Domain (GTP/GDP-binding, ~14 kDa) and Beta Barrel 2 Domain (~12 kDa). The inactive GTP-bound enzyme is present in a closed conformation, which upon activation by Ca2+ and substrate binding opens like a pocket knife resulting in a longitudinal open conformation (see figure). Figure 4: closed (left) and open (right) conformation of human tissue transglutaminase Blue: Beta Sheet Domain Yellow: catalytic Core Domain Pink: Beta Barrel 1 Domain Green: Beta Barrel 2 Domain

Host

Rabbit

Immunogen

Human tissue transglutaminase (full length protein with N-terminal hexahistidin-tag) recombinantly produced in insect cells

Specificity

Specificity of A014 was determined with human transglutaminases (TG1 – TG7, FXIII) and TG2 of different species. Cross-reactivity of A014 to other rec. human transglutaminases Figure 1: Equal amounts of human transglutaminases (TG1 – TG7, FXIII) were analysed by SDS-PAGE-Gel (staining with coomassie brilliant blue, left) and western blot (right, A014 diluted 1:2,000 in TBST; 2nd AB: Anti-Rabbit IgG - Alkaline Phosphatase antibody diluted 1:10,000 in TBST). The western blot reveals that A014 has no cross reactivity to TG1, TG6 and TG7 and very slight cross reactivity to TG3, TG4, TG5 and FXIII. Cross-reactivity of A014 to rec. TG2 from various species Figure 2: Equal amounts (1 µg and 0.3 µp protein per lane respectively) of transglutaminase 2 from various species (human, guinea pig (gp), rat and mouse) were analysed by SDS-PAGE Gel (staining with coomassie brilliant blue, left) and western blot (right, A014 diluted 1:2,000 in TBST; 2nd AB: Anti-Rabbit IgG - Alkaline Phosphatase antibody diluted 1:10,000 in TBST). The western blot shows that A014 also recognizes TG2 from various species, although at a lower signal intensity. Note: A014 also detects dog TG2.

Formulation

The antibody is lyophilized from PBS, 2 mg/mL human serum albumin, azide free.

Application

Western-Blotting, immuno-fluorescence, immuno-histochemistry Note: For FACS antibody A028 (FITC-labeled A014) is recommended.

Working dilutions

Optimal dilutions should be determined by the end user. For Western-Blotting: 1 / 500 to 1 / 5,000 should be suitable For immuno-fluorescence and immuno-histochemistry 1 / 100 to 1 / 500 is recommended.

Storage

Stable for a minimum of 5 years at –80°C as lyophilized powder.

Reference(s)

Dulay et al., Biosens. Bioelectron. 2011, 26:3852-6

Note

INTENDED FOR RESEARCH USE ONLY, NOT FOR USE IN HUMAN, THERAPEUTIC OR DIAGNOSTIC APPLICATIONS.

Related Products

A028: FITC-labeled polyclonal antibody to human tissue transglutaminase (TG2)
A053: Biotinylated polyclonal antibody to human tissue transglutaminase (TG2)
A033: Monoclonal antibody to human TG2 (Catalytic Domain, clone XTG17)
T002: Human tissue transglutaminase