AMBP (Human) ELISA Kit

Sample

Cell Culture Supernatant, Cell Lysate, CSF, Milk, Plasma, Saliva, Serum, Urine

Species Reactivity

Human

Detection Method

sELISA

Intended Use

AMBP (Human) ELISA Kit is a sandwich enzyme immunoassay for the quantitative measurement of human alpha1-microglobulin.

Contents of Kit

1. Human alpha-1-Microglobulin Microplate: A 96-well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody against human alpha-1-microglobulin. 96 (8×12) wells
2. Sealing Tapes: Pressure Sensitive sealing tapes that can be cut to fit the format of the individual assay. 3 slices
3. Human alpha-1-Microglobulin Standard: Human alpha1-microglobulin in a buffered protein base (lyophilized). 14 ng
4. Biotinylated Human alpha-1-Microglobulin Antibody (50×): A 50-fold concentrated biotinylated polyclonal antibody against human alpha-1-microglobulin. 120 μL
5. EIA Diluent Concentrate (10×): A 10-fold concentrated buffered protein base. 30 mL
6. Wash Buffer Concentrate (20×): A 20-fold concentrated buffered surfactant. 30 mL × 2
7. SP Conjugate (100×): A 100-fold concentrate. 80 μL
8. Chromogen Substrate (1×): A stabilized peroxidase chromogen substrate tetramethylbenzidine. 7 mL
9. Stop Solution (1×): A 0.5 N hydrochloric acid solution to stop the chromogen substrate reaction. 11 mL

Storage

1. Upon arrival, immediately store components of the kit at recommended temperatures up to the expiration date.
2. Store SP Conjugate and Biotinylated Antibody at -20°C.
3. Store Microplate, Diluent Concentrate (10×), Wash Buffer, Stop Solution, and Chromogen Substrate at 2-8°C.
4. Unused microplate wells may be returned to the foil pouch with the desiccant packs and resealed. May be stored for up to 30 days in a vacuum desiccator.
5. Store Standard at 2-8°C before reconstituting with Diluent and at -20°C after reconstituting with Diluent

Precision

Intra-assay precision was determined by testing three plasma samples twenty times in one assay.
Inter-assay precision was determined by testing three plasma samples in twenty assays.

Detection Limit

The minimum detectable dose of human alpha-1-microglobulin as calculated by 2SD from the mean of a zero standard was established to be 0.16 ng/mL.

General Description

Alpha-1-microglobulin (A1M), also called protein HC, is a tubular plasma and tissue protein that belongs to the lipocalin transport protein superfamily for small hydrophobic molecules. It contains 184 amino acids and has a molecular weight of 26 kDa. Mature A1M and bikunin (urinary trypsin inhibitor) result from a common precursor. A1M is found in blood, both in free form and complex-bound to immunoglobulin A (IgA). It is involved in inflammatory and detoxification processes caused by immune system activation and extracellular heme catabolism. While increased excretion is detected in urine or serum shortly after tubular injury, A1M may predict acute kidney injury and the need for renal replacement therapy. Urinary A1M is useful for the early detection of nephropathy in type 2 diabetic subjects.

Standard Curve

The typical data is provided for reference only. Individual laboratory means may vary from the values listed. Variations between laboratories may be cause by technique differences.

The curve is provided for illustration only. A standard curve should be generated each time the assay is performed.

Reference Value
Normal human alpha-1-microglobulin plasma levels range from 31.5 - 58.5 μg/mL.
Plasma and serum samples from healthy adults were tested (n=40). On average, human alpha-1-microglobulin level was 44.3 μg/mL.