Albumin ELISA Kit

Produktnummer ABIN612654

Produktdetails Albumin ELISA Kit

Target: Albumin (ALB)

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Untere Nachweisgrenze: 3 ng/mL

Applikation: ELISA

Verwendungszweck: The AssayMax Human Albumin ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for detection of human albumin in urine, saliva, milk, and cell culture supernatants

Marke: AssayMax

Proben: Cell Culture Supernatant

Analytische Methode: Quantitative

Spezifität: 10% FBS in culture media will not affect the assay.

Bestandteile: Human Albumin Microplate: A 96-well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody against human albumin. Sealing Tapes: Each kit contains 3 pre-cut, pressure-sensitive sealing tapes that can be cut to fit the format of the individual assay. Human Albumin Standard: Human albumin in a buffered protein base (800 ng, lyophilized). Biotinylated Human Albumin Antibody (100x): A 100-fold concentrated biotinylated polyclonal antibody against human albumin (80µl). 1 MIx Diluent Concentrate (10x): A 10-fold concentrated buffered protein base (30 ml). Wash Buffer Concentrate (20x): A 20-fold concentrated buffered surfactant (30 ml, 2 bottles). Streptavidin-Peroxidase Conjugate (SP Conjugate): A 100-fold concentrate (80µl). Chromogen Substrate: A ready-to-use stabilized peroxidase chromogen substrate tetramethylbenzidine (8 ml). Stop Solution: A 0.5 N hydrochloric acid to stop the chromogen substrate reaction (12 ml).

Benötigtes Material: Microplate reader capable of measuring absorbance at 450 nm. Pipettes (1-20 µL, 20-200 µL, 200-1000µLand multiple channel). Deionized or distilled reagent grade water

Anwendungsinformationen

Probenmenge: 50 μL

Testdauer: < 3 h

Plattentyp: Pre-coated

Protokoll: This assay employs a quantitative sandwich enzyme immunoassay technique that measures human albumin in less than 3 hours. A polyclonal antibody specific for human albumin has been pre-coated onto a 96-well microplate with removable strips. Albumin in standards and samples is sandwiched by the immobilized polyclonal antibody and biotinylated polyclonal antibody specific for human albumin, which is recognized by a streptavidin-peroxidase conjugate. All unbound material is then washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.

Aufbereitung der Reagenzien:

Freshly dilute all reagents and bring all reagents to room temperature before use. If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. MIx Diluent Concentrate (10x): Dilute the MIx Diluent 1:10 with reagent grade water. Store for up to 1 month at 2-8°C. Standard Curve: Reconstitute the 800 ng of Albumin Standard with 4 ml of MIx Diluent to generate a standard solution of 200 ng/ml. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. Prepare duplicate or triplicate standard points by 2 serially diluting the standard solution (200 ng/ml) 1:2 with equal volume of MIx Diluent to produce 100, 50, 25 12.5, 6.25, and 3.125 ng/ml solutions. MIx Diluent serves as the zero standard (0 ng/ml). Any remaining solution should be frozen at -20°C. Standard Point Dilution [H. Albumin] (ng/ml) P1 Standard (200 ng/ml) 200.000 P2 1 part P1 + 1 part MIx Diluent 100.000 P3 1 part P2 + 1 part MIx Diluent 50.000 P4 1 part P3 + 1 part MIx Diluent 25.000 P5 1 part P4 + 1 part MIx Diluent 12.500 P6 1 part P5 + 1 part MIx Diluent 6.250 P7 1 part P6 + 1 part MIx Diluent 3.125 P8 MIx Diluent 0.000 Biotinylated Human Albumin Antibody (100x): Spin down the antibody briefly and dilute the desired amount of the antibody 1:100 with MIx Diluent. Any remaining solution should be frozen at -20°C. Wash Buffer Concentrate (20x): Dilute the Wash Buffer Concentrate 1:20 with reagent grade water. SP Conjugate (100x): Spin down the SP Conjugate briefly and dilute the desired amount of the conjugate 1:100 with MIx Diluent. Any remaining solution should be frozen at -20°C.

Probennahme: Cell Culture Supernatants: Centrifuge cell culture media at 3000 x g for 10 minutes to remove debris. Collect supernatants and assay. Store samples at -20°C or below. Avoid repeated freeze-thaw cycles. Urine: Collect urine using sample pot. Centrifuge samples at 600 x g for 10 minutes and assay. Urine dilution is suggested at 1:200 into MIx Diluent, however, the user should determine the optimal dilution factor. Store samples at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles Saliva: Collect saliva using sample tube. Centrifuge samples at 600 x g for 10 minutes and assay. Saliva dilution is suggested at 1:200 into MIx Diluent, however, the user should determine the optimal dilution factor. Store samples at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles. Milk: Collect milk using sample tube. Centrifuge samples at 800 x g for 10 minutes and assay. Milk dilution is suggested at 1:6000 into MIx Diluent, however, the user should determine the optimal dilution factor. The undiluted samples can be stored at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles.

Testdurchführung:

Prepare all reagents, working standards and samples as instructed. Bring all reagents to room temperature before use. The assay is performed at room temperature (20 - 30 °C). Remove excess microplate strips from the plate frame and return them immediately to the foil pouch with desiccant inside. Reseal the pouch securely to minimize exposure to water vapor and store in a vacuum desiccator. Add 50 µL of standard or sample per well. Cover wells with a sealing tape and incubate for one hour. Start the timer after the last sample addition. Wash five times with 200 µL of Wash Buffer manually. Invert the plate each time and decant the contents, hit it 4-5 times on absorbent paper towel to completely remove the liquid. If using a machine wash six times with 300 µL of Wash Buffer and then invert the plate, decant the contents, hit it 4-5 times on absorbent paper towel to completely remove the liquid. Add 50 µL of Biotinylated Human Albumin Antibody to each well and incubate for 30 minutes. Wash a microplate as described above. Add 50 µL of Streptavidin-Peroxidase Conjugate to each well and incubate for 30 minutes. Turn on the microplate reader and set up the program in advance. Wash a microplate as described above. Add 50 µL of Chromogen Substrate per well and incubate for about 15 minutes or till the optimal blue color density develops. Gently tap plate to ensure thorough mixing and break the bubbles in the well with pipette tip. Add 50 µL of Stop Solution to each well. The color will change from blue to yellow. 3 Read the absorbance on a microplate reader at a wavelength of 450 nm immediately. If wavelength correction is available, subtract readings at 570 nm from those at 450 nm to correct optical imperfections. Otherwise, read the plate at 450 nm only. Please note that some unstable black particles may be generated at high concentration points after stopping the reaction for about 10 minutes, which will reduce the readings.

Ergebnisberechnung:

Calculate the mean value of the triplicate for each standard and sample. To generate a Standard Curve from the initial reaction time, plot the graph using the standard concentrations on the x-axis and the corresponding mean 405 nm absorbance or change in absorbance per minute (A/min) on the y-axis after subtracting the background. The best-fit line can be determined by regression analysis of the linear portion of the curve. Determine the unknown sample concentration from the Standard Curve and multiply the value by the dilution factor of 5. Standard Curve The curve is provided for illustration only. A standard curve should be generated each time the assay is performed.

Testpräzision: Intra-assay and inter-assay coefficients of variation were 4.8% and 7.2% respectively.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: The kit should not be used beyond the expiration date.

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung: Store kit at 2-8°C or -20°C upon arrival up to the expiration date. Opened MIx Diluent may be stored for up to 1 month at 2-8°C. Store reconstituted reagents at -20°C or below. Opened unused strip wells may return to the foil pouch with the desiccant pack, reseal along zip-seal. May be stored for up to 1 month in a vacuum desiccator.

Referenzen für Albumin ELISA Kit (ABIN612654)

Huang, Cui, Peng, Wu, Han, Dong: "Preparation of three-dimensional macroporous chitosan-gelatin B microspheres and HepG2-cell culture." in: Journal of tissue engineering and regenerative medicine, (2014) (PubMed).

Wiegand, Hewitt, Merk, Reisinger: "Dermal xenobiotic metabolism: a comparison between native human skin, four in vitro skin test systems and a liver system." in: Skin pharmacology and physiology, Vol. 27, Issue 5, pp. 263-75, (2014) (PubMed).

Details zu Albumin

Target: Albumin (ALB)

Andere Bezeichnung: Albumin (ALB Produkte)

Synonyme: PRO0883 ELISA Kit, PRO0903 ELISA Kit, PRO1341 ELISA Kit, ALB ELISA Kit, CSA ELISA Kit, Alb-1 ELISA Kit, Alb1 ELISA Kit, Albza ELISA Kit, LOC100136344 ELISA Kit, alb-a ELISA Kit, alb-b ELISA Kit, albumin ELISA Kit, serum albumin 1 ELISA Kit, albumin S homeolog ELISA Kit, ALB ELISA Kit, Alb ELISA Kit, LOC100136344 ELISA Kit, alb.S ELISA Kit

Hintergrund: Albumin is a serum hepatic protein, the most abundant protein in serum and contributes to the maintenance of oncotic pressure as well as to transport of hydrophobic molecules. Serum albumin level has been linked in clinical practice to several diseases. Low albumin levels can suggest liver , kidney disease , inflammation , shock , and malnutrition. On the other hand, high albumin levels usually reflect dehydration.

Pathways: Lipid Metabolism