Albuwell M: A murine microalbuminuria ELISA.

Intended Use: Albuwell M is an enzyme linked immunosorbent assay (ELISA) for the quantitative determination of albumin in mouse urine.

Kit Contents: Your Albuwell M kit should contain the following items:
1. 2 Albuwell M Test Plates
2. 2 NHEBSA (Diluent)
3. Mouse Serum Albumin (MSA) Standard
4. 2 Anti-mouse Albumin Ab-HRP Conjugate
5. 2 Color Developer
6. 2 Color Stopper
7. Instructions

Technical Background: Albuwell M is an in-vitro tool for assessing kidney function in mice. It is simple to perform and highly specific for mouse albumin. It is a competitive antibody capture ELISA completed in a direct mode. To that end, the anti-albumin antibody is conjugated to horseradish peroxidase (anti-mosue Albumin Ab-HRP), i.e. directly labeled.

To complete the assay, sample and anti-mouse albumin Ab-HRP are added to mouse albumin-coated well. The antibody interacts and binds with the albumin immobilized onto the stationary phase or with that in the fluid phase, hence the notion of competitive binding. Washing removes unbound Ab-HRP-Albumin and other reactants of the fluid phase from the well. Only the antibody-conjugate that was bound to the albumin of the stationary phase remains, and this is detected using Tetramethylbenzidine (TMB) in a chromogenic reaction. The reaction is stopped with acid, and absorbance is measured at 450 nm. Absorbance is inversely proportional to the logarithm of albumin concentration in the fluid phase.

Specimen Collection and Storage: Collect samples without preservative, and clarify them by centrifugation if necessary. 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.

MSA Standard, NHEBSA and anti-mouse Albumin Ab-HRP Conjugate preparations contain 0.05% Proclin 300 (active components isothiazolones) as preservative. Color Stopper contains dilute (2.0 N) sulfuric acid. Albuwell M plates are precoated and ready to use. All kit reagents are supplied in ready to use liquid form.