ELISA Substrates for alkaline phosphatase (AP) - slow

Cat. No.: E491324

GRADE & PURITY slow

Storage

Store at 2-8°C,Protected from light

Shipped In

Wet ice

Overview

1、Product attribute

Shelf life: 24 months

Reaction time：long (up to 45 minutes) at 20-37°C

Lot-to-lot variation：<10%

Boiling point : 100℃

pH-Value (at 20 °C): 9.0-9.8   

Density (20℃) : 1.0302 g/cm³

Water solubility: easily soluble

Appearance: colourless to light yellow liquid

Odour: odourless

Incubation temperature: 20-37 °C

Light sensitive
Heat sensitive

2、Requirements for storage rooms and vessels

1.Keep container tightly closed.

2.Keep cool. protected from light

3. Do not store together with: Oxidizing agent. 

4. Contaminated or leaked out substrate solution from damaged bottles should not be used anymore and has to be destroyed.

5. Use isolated containers with some cool bags for transport.

6. Spontaneous decay will increase the background. If stored at room temperature, the velocity of the decay will increase. Thus, both storage and transport at room temperature should be avoided. Nevertheless, the activity of the solution is not affected by storage at room temperature. The solution still works beyond the expiry date, but some applications, especially those including visual evaluation, may be hampered by increased background.

3、Effective Components and Principle of Function

In different buffer solutions (pH = 9.5), with supplementation if required, the effective component

para nitrophenyl phosphate (pNPP) is dissolved.

 Alkaline Phosphatase transfers the phosphate residue to an acceptor. Under alkaline conditions a yellow colour occurs, resulting from the formed nitrophenol.

4、Biosafety information

This mixture is not classified as hazardous in accordance with Regulation (EC) No 1272/2008；

5、Advantage

1. Signal yield comparable to competitor ready-to-use solutions

2. Broad measurement range

3. Very low background signals

4. Very low blank drift during long-term storage (<0.15 AU within 24 months)

5. High colour stability after reaction stop with this product and other commonly used stopping solutions

6、Instruction for usage

For bottling consider the following instructions:

•  Work in a dust free and darkened room.

• Keep the solution as cool as possible.

• Avoid contact of the solutions with any metal parts

• Clean all instruments and vessels very extensively.

• Wear powder-free gloves during bottling.

• Close the bottles immediately to minimize the influence of light and dust.

• Use clean bottles that are impermeable to light made from HDPE or PP.

7、 General Instructions for the Use in Blotting Systems 

Only qualified laboratory staff, who are familiar with the basics of immunological methods, are allowed to use these solutions.

The substrate solutions can be used in qualitative and quantitative ELISA procedures.

When using 96-well microtiter plates, adding 100 μL of substrate per well after incubation and washing is recommended. After substrate incubation the reaction can be stopped and the photometric measurement can be carried out. Using higher incubation temperatures (37° C) may shorten the incubation time. 

The reaction can be stopped by using the special developed solution stop. The use of other commercially available stop solutions cannot safely exclude a further increase of the signal. Addition of a stopping solution does not change the general shape of the spectrum. The unstopped and the stopped solution should be measured at 405 nm and the background correction: should be measured at 620 nm.

Specifications

Specifications & Purity

slow

Storage

Store at 2-8°C, Protected from light

Shipped In

Wet ice

This product requires cold chain shipping. Ground and other economy services are not available.

Names and Identifiers

• PH: 9.0-9.8

Chemical and Physical Properties

• Solubility: Easily soluble in water
• Sensitivity: Light & heat sensitive