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Albuwell O (Sheep Albumin ELISA)

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The Albuwell O kit contains all of the components and reagents you need to assay sheep albumin in up to 80 duplicate urine or cell free liquid sameples (plasma, serum, cell culture supernat, etc....)

Intended Use:  Albuwell O is an enzyme linked immunosorbent assay (ELISA) for the quantitative determination of albumin in sheep urine.  It is for research purposes, and is not intended for diagnostic use.

Technical Background:  Albuwell O is for in vitro use as a research tool for assessing kidney function in sheep.  It is simple to perform and highly specific for sheep albumin.  Albuwell O is a conventional indirect competitive ELISA that recognizes antigen (albumin) in test samples.

To complete the assay, sample and rabbit anti-ovine albumin antibody are added to an albumin-coated well.  The antibody interacts and binds with the albumin immobilized to the stationary phase or with that in the fluid phase, hence the notion of competitive binding.  A subsequent reaction with antirabbit -HRP conjugate labels the probe with enzyme.  After washing, only the antibodyconjugate that bound to the albumin of the stationary phase remains in the well, and this is detected using a chromogenic reaction.  Color intensity in Albuwell O is inversely proportional to the logarithm of albumin in the fluid phase.

Specimen Collection and Storage:  Collect samples without preservative, and render them particulate free by centrifugation.  Store clarified urine at 4oC for up to 1 week or at 60oC for up to 2 months.  Prior to assay, allow the samples to come to room temperature.  Do not apply heat to thaw frozen samples.  Clarify samples by centrifugation if necessary.

Kit Contents:  Your Albuwell O kit should contain the following items:

  1. 2 Albuwell O Test Plates
  2. 2 NHE-BSA Diluent
  3. 2 Ovine Serum Albumin Standard
  4. 2 Rabbit Anti-ovine Albumin Antibody
  5. 2 Anti-rabbit HRP Conjugate
  6. 2 Color Developer
  7. 2 Color Stopper
  8. Instructions

Ovine Serum Albumin (OSA) Standard, NHEBSA Diluent, Rabbit anti-ovine Albumin Antibody and Anti-rabbit HRP Conjugate preparations contain 0.05% Proclin 300 (active components isothiazolones) as preservative.  Color Stopper contains dilute (2.0 N) sulfuric acid. 

Albuwell O plates are precoated and ready to use.  All kit reagents are supplied in ready to use liquid form.  A provision to wash the plates should be made, an EIA Wash Buffer with composition: 0.15 M NaCl, 0.01 M triethanolamine (pH 6.8), 0.05% Tween 20 and 0.05 % Proclin 300 (preservative may be omitted if the buffer is freshly prepared) is recommended.

Micropipettors capable of delivering 10, 50, 100 and 120 uL are required.  Multi-channel pipettors capable of delivering 50 and 100 uL are recommended.  In addition, small test tubes are required to complete dilutions (microfuge tubes work well in this application).  Finally, a microplate reader equipped to determine absorbance at 450 nm is required.

Assay Procedure: Allow reagents and samples to come to room temperature before running the assay.


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