PEA ELISA Kit (Pseudomonas Exotoxin A)

INTENDED USE

Human PEA ELISA kit can be used for analysing levels of pseudomonas exotoxin A (PEA) present in human samples of serum, tissue homogenates, plasma and other biological fluids.

INTRODUCTION

Pseudomonas exotoxin A (PEA) is a 638 amino acid pro-protein that is subject of extensive study due its potential to be used as a therapeutic agent. This pro-protein is converted to the mature toxin that is 613 amino acids in length and then secreted. The mature protein consists of three main functional domains a catalytic domain, a receptor binding domain and a translocation domain. These well define structural division make it an ideal candidate for developing other chimeric toxins.

There is evidence to indicate that PEA is like other bacterial toxin, where it has identical mechanism of action similar to diphtheria toxin. For example, both can induce apoptosis in mast cells found in humans using the mediated mechanism of caspase 3 and caspase 6. The presence of these similar toxins indicates a modular recombination process occurring during this toxin evolution. This is the same enzyme targeted by tetrodotoxin. The toxicity of pseudomonas exotoxin A on “Xenopus laevis” cells was tested using the ATPase assay system which revealed a significant inhibition of cellular ATP production in these cells following exposure to 10 micromolar doses of PEA.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti-PEA antibody.
• Standards: Lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

TYPICAL RESULTS

For this PEA ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 31.25, 62.5, 125, 250, 500, 1000, 2000 pg/ml.
Reactivity: Human
Sensitivity: 18.75 pg/ml
Range: 31.25 – 2000 pg/ml
Principle: Sandwich
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for PEA, no cross reactivity or interference between PEA and analogues was detected.
– Recovery: Serum (88 – 102%), EDTA Plasma (85 – 104%), Heparin Plasma (85 – 103%).
– Linearity: Serum (86 – 102%), EDTA Plasma (86 – 94%), Heparin Plasma (81 – 99%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

1. The immunotoxin activity of exotoxin A is sensitive to domain modifications. Int J Biol Macromol. (2019) 134: 1120-1131. Amiri Tehranizadeh Z., et al.
2. Genetic deficiency in low density lipoprotein receptor-related protein confers cellular resistance to Pseudomonas exotoxin_A. Evidence that this protein is required for uptake and degradation of multiple ligands. Cell Sci. (1994) 107 (Pt 3): 719-26. Willnow T.E. and Herz J.J.
3. Calmodulin antagonists sensitize cells to pseudomonas toxin. J Cell Physiol. (1984) 119 (1): 15-22. Sundan A., et al.

ADDITIONAL INFORMATION

• Full Name: PEA ELISA Kit (Pseudomonas Exotoxin A)
• Reactivity: Human
• Sample Type: Serum, Biological Fluids, Plasma, Tissue Homogenates
• Sensitivity: 18.75 pg/ml

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