Pancreatic Elastase ELISA Kit

$830.00

The Eagle Biosciences Pancreatic Elastase ELISA Assay Kit is intended for the quantitative determination of human pancreatic elastase in stool and fecal samples. The Pancreatic Elastase ELISA is for research use only and should not be used in therapeutic procedures.

Pancreatic Elastase ELISA Assay Kit

Pancreatic Elastase ELISA Kit manufactured in Germany by Immundiagnostik

Size: 1×96 wells
Sensitivity: 0.66 µg/mL
Incubation Time: 2 hours
Sample Type: Stool
Sample Size: 15 mg
For Research Use Only

Controls Included
Reference range:
Reference range in stool samples:
1 g stool is equivalent to 1 ml.
> 200µg/ml, normal value
100 – 200µg/ml, slight to moderate exocrine pancreatic insufficiency
< 100µg/ml, exocrine pancreatic insufficiency
We recommend each laboratory to establish its own reference concentration range.
Specificity:
No cross reactivity to the following proteins was observed:
• Pancreatic lipase
• Chymotrypsin
• Pankreatic amylase
• Pancreatin
• Calprotectin


Assay Principle

The Eagle Biosciences Pancreatic Elastase ELISA Kit is intended for the quantitative determination of pancreatic elastase in stool. In a first incubation step, the pancreatic elastase in the samples is bound to monoclonal antibodies, immobilized to the surface of the microtiter wells. To remove all unbound substances, a washing step is carried out. In a second incubation step, a peroxidase-labeled conjugate (mouse anti pancreatic elastase) is added which recognizes specifically the bound pancreatic elastase. After another washing step to remove all unbound substances, the solid phase is incubated with the substrate, tetramethyl-benzidine (TMB), which reacts with the peroxidase. An acidic stop solution is added to stop the reaction. The color changes from blue to yellow. The intensity of the yellow color is directly proportional to the concentration of pancreatic elastase. A dose response curve of absorbance unit (optical density, OD at 450 nm) vs. concentration is generated using the values obtained from the standards. Pancreatic elastase present in the patient samples is determined directly from this curve.


Related Products

Pancreatic Amylase ELISA Assay Kit
PMN-Elastase (Stool) ELISA Assay Kit
PMN Elastase ELISA Assay Kit

Additional Information

Assay Background


Pancreatic elastase is an anionic endoprotease of the serine protease family with a molecular weight of 26 kDa. Together with other digestive enzymes it is synthesized as an inactive pro-enzyme in the acinar cells of the pancreas and is secreted into the duodenum. After its activation, pancreas elastase cleaves peptides after neutral amino acids. Pancreas elastase is mainly bound to bile salts during intestinal passage and is not degraded. In human feces it is 5–6 fold more concentrated than in pancreatic juice. The stool concentration reflects the secretory capacity of the pancreas.

Assay Procedure


  1. Add 100 µl of STD/SAMPLE/CTRL (standard / sample / controls) into respective well.
  2. Cover the strips and incubate for 30 min at room temperature (15-30 °C) on a horizontal shaker.
  3. Discard the contents of each well and wash 5 times with 250 µl wash buffer. After the final washing step, the inverted microtiter plate should be firmly tapped on absorbent paper.
  4. Add 100 µl conjugate to each well.
  5. Cover the strips and incubate for 30 min at room temperature (15-30 °C) on a horizontal shaker.
  6. Discard the contents of each well and wash 5 times with 250 µl wash buffer. After the final washing step, the inverted microtiter plate should be firmly tapped on absorbent paper.
  7. Add 100 µl SUB (TMB substrate) in each well.
  8. Incubate for 10-20 minutes at room temperature (15-30 °C) in the dark.
  9. Add 100 µl STOP (ELISA stop solution) and mix well.
  10. Determine absorption immediately with an ELISA reader at 450 nm against 620 nm (or 690 nm) as a reference. If no reference wavelength is available, read only at 450 nm. If the extinction of the highest standard exceeds the range of the photometer, absorption must be measured immediately at 405 nm against 620 nm as a reference.

Manual

Product Manual


Publications

References


  • Nandhakumar, N. & Green, M.R., 2010. Interpretations: How to use faecal elastase testing. Archives of disease in childhood. Education and practice edition, 95(4), pp.119–23.
  • Whitcomb, D.C. & Lowe, M.E., 2007. Human pancreatic digestive enzymes. Diges- tive diseases and sciences, 52(1), pp.1–17.
  • Dominici, R. & Franzini, C., 2002. Fecal elastase-1 as a test for pancreatic function: a review. Clinical chemistry and laboratory medicine : CCLM / FESCC, 40(4), pp.325–32.
  • Löser, C., Möllgaard, A. & Fölsch, U.R., 1996. Faecal elastase 1: a novel, highly sensi- tive, and specific tubeless pancreatic function test. Gut, 39(4), pp.580–6.
  • Stein, J. et al., 1996. Immunoreactive elastase I: clinical evaluation of a new nonin- vasive test of pancreatic function. Clinical chemistry, 42(2), pp.222–6.