UBA3 ELISA Kit

Produktnummer ABIN2949769

Das Human UBA3 ELISA Kit (ABIN2949769) ist ein Colorimetric ELISA Kit zur Detektion von Human UBA3.

Kurzübersicht für UBA3 ELISA Kit (ABIN2949769)

Target: UBA3 (Ubiquitin-Like Modifier Activating Enzyme 3 (UBA3))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 0.156 ng/mL - 10 ng/mL

Applikation: ELISA

Proben: Cell Lysate, Tissue Homogenate

Produktdetails UBA3 ELISA Kit

Untere Nachweisgrenze: 0.156 ng/mL

Verwendungszweck: This kit is based on sandwich enzyme-linked immuno-sorbent assay technology.

Analytische Methode: Quantitative

Sensitivität: < 0.05 ng/mL

Bestandteile: The kit components listed are for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product.

• Pre-coated 96-Well Microplate
• Standard
• Standard Diluent Buffer
• Wash Buffer
• Detection Reagent A
• Detection Reagent B
• Diluent A
• Diluent B
• TMB Substrate
• Stop Solution
• Plate Sealer

Benötigtes Material:

• 37 °C incubator
• Multi and single channel pipettes and sterile pipette tips
• Squirt bottle or automated microplate washer
• 1.5 mL tubes
• Distilled water
• Absorbent filter papers
• 100 mL and 1 liter graduated cylinders
• Microplate reader (wavelength: 450 nm)
• ELISA Shaker

Anwendungsinformationen

Probenmenge: 100 µL

Plattentyp: Pre-coated

Protokoll: This kit is based on sandwich enzyme-linked immuno-sorbent assay technology. An antibody is pre-coated onto a 96-well plate. Standards, test samples, and biotin-conjugated reagent are added to the wells and incubated. The HRP-conjugated reagent is then added, and the whole plate is incubated. Unbound conjugates are removed using wash buffer at each stage. TMB substrate is used to quantify the HRP enzymatic reaction. After TMB substrate is added, only wells that contain sufficient UBA3 will produce a blue coloured product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow colour is proportional to the UBA3 amount bound on the plate. The Optical Density (OD) is measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of UBA3 can be calculated.

Aufbereitung der Reagenzien:

This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product.

• 1) Standard: Prepare the standard with the recommended volume of Standard Diluent Buffer, to make the standard solution. Then use the Standard Diluent buffer to carry out serial dilutions of the standard solution, as instructed in the Protocol.

• 2) Wash Buffer: Dilute the concentrated Wash Buffer with distilled water, as instructed in the Protocol.

• 3) Detection Reagent Preparation: Calculate the total volume of working solution required. Dilute Detection Reagent A and Detection Reagent B with Diluent A and Diluent B, respectively, at 1:100.

Testdurchführung:

This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product.

• 1) Set standard, test samples and control wells.

• 2) Aliquot 100 μL of diluted standard into the standard wells.

• 3) Aliquot 100 μL of Standard Diluent buffer into control (zero) well.

• 4) Aliquot 100 μL of diluted samples into the sample wells. Incubate for 1 hr at 37 °C.

• 5) Aliquot 100 μL of Detection Reagent A to each well. Incubate for 1 hr at 37 °C.

• 6) Wash 3 times.

• 7) Aliquot 100 μL of Detection Reagent B to each well. Incubate for 30 mins at 37 °C.

• 8) Wash 5 times.

• 9) Aliquot 90 μL of TMB Substrate to each well. Incubate for 10-20 mins at 37 °C.

• 10) Aliquot 50 μL of Stop Solution.

• 11) Measure the OD at 450 nm.

Ergebnisberechnung:

For calculation, average the O.D.450 duplicate readings for each reference standard and each sample and substract the average control (zero) O.D.450 reading. The standard curve can be plotted as the relative O.D.450 of each reference standard solution (Y) vs. the respective concentration of each standard solution (X). The UBA3 concentration of the samples can be interpolated from the standard curve.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: 4 °C

Informationen zur Lagerung: Shipped at 4°C. Upon receipt, store the kit according to the storage instruction in the kit's manual.The stability of the kit is determined by the rate of activity loss. The loss rate is less than 5% within the expiration date under appropriate storage conditions. To minimize performance fluctuations, operation procedures and lab conditions should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same user throughout.

Haltbarkeit: 6 months

Details zu UBA3

Target: UBA3 (Ubiquitin-Like Modifier Activating Enzyme 3 (UBA3))

Andere Bezeichnung: NEDD8-activating enzyme E1 catalytic subunit

UniProt: Q8TBC4