Lowry Protein Quantification Assay Kit

Produktnummer ABIN3172698

Produktdetails

Nachweismethode: Colorimetric

Detektionsbereich: 1-1500 μg/mL

Untere Nachweisgrenze: 1 μg/mL

Applikation: Quantification (Q)

Verwendungszweck: The kit is designed for protein quantification in biological samples.

Proben: Plasma, Tissue Homogenate, Urine, Serum, Cell Lysate

Analytische Methode: Quantitative

Spezifität: Specific for protein quantification.

Produktmerkmale: Lowry Protein Quantification Assay is based on Lowry method1, first described in 1951. The method relies on two different reactions. The first is the formation of a copper ion complex with amide bonds, forming reduced copper in alkaline solutions. This is

Bestandteile: Plate and the reagents neccesary to perform the assay.

Benötigtes Material: Pipettes, reaction tubes, plate reader, centrifuge.

Anwendungsinformationen

Kommentare:

1ml (microassay)/ 40μL (microplate)

Testdauer: 1 h

Plattentyp: Uncoated

Aufbereitung der Reagenzien:

Lowry Assay Mix: Mix Lowry Reagent A with Lowry Reagent B in a ratio 1:100. Prepare fresh daily.
Lowry Working Solution: Dilute Lowry Reagent C with an equal volume of water to prepare the desired volume. Prepare diluted reagent fresh daily.

Testdurchführung:

MICROASSAY: 1. Pipette1 mL of each standard or unknown sample solution into separate clean test tubes. Refer to the Table 2 as a guide for diluting the protein standard (see kit book let). For the diluents, use the same buffer as in the samples. 2.To each tube, add 5 mL of freshly prepared Lowry assay mix and thoroughly vortex. 3.Incubate tubes 10 min at room temperature. 4.Add 0.5 mL of Lowry Working Solution to each tube and vortex immediately. 5.Incubate 30 min at room temperature. 6.Vortex the tubes, zero the spectrophotometer with the blank, and measure the absorbance at 660 nm.
MICROPLATE: 1. Pipette 40 μL of each standard and unknown sample replicate into a microplate well. Refer to the Table 3 as a guide for diluting the protein standard (see kit book let). For the diluents, use the same buffer as in the samples. 2.Add 200 μL of freshly prepared Lowry Assay Mix and immediately mix the microplate for 30 seconds. 3.Incubate microplate exactly 10 min at room temperature. 4.Add 20 μL of freshly prepared Lowry Working Solution to each well and immediately mix the microplate for 30 seconds. 5.Incubate 30 min at room temperature. Measure the absorbance at 660 nm on a plate reader.

Ergebnisberechnung:

1.If the spectrophotometer or microplate reader was not zeroed with the blank, then average the blank values and substract the average blank value from the standard and unknown sample values. 2.Create a standard curve by plotting A660nm (y-axis) vs BSA standard (μg) (x-axis). Determine the unknown sample concentration using the standard curve . 3.The level of detection of the assay is lower for the microplate assay when compared with the microassay due to a shorter light path used in the microplate reader. 4.Standard curve example for microplate assay procedure is shown in Figure 2 (see Images). A sample Lowry protein assay standard curve produced using BSA at triplicate points in a range of 5 to 280 μg is shown in Figure 3 (see images). The data are fit with a polynomic regression.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Lagerung: 4 °C