QUANTI-Star™: ACP5 Detection Reagent

Colorimetric-based assay reagent for the detection of acid phosphatase 5 (ACP5)

QUANTI-Star™ is a liquid formulation of a colorimetric enzyme assay. It has been developed to determine the release of acid phosphatase 5 (ACP5) in a biological sample, such as cell culture supernatants.

Description

ACP5, also known as tartrate-resistant acid phosphatase (TRAP or TRAPase) is a metalloprotein enzyme, usually located in the lysosome. InvivoGen has developed a secreted (s) version of ACP5 (sACP5) that can be used as a reporter gene.

The level of sACP5 activity in the cell culture supernatant is easily quantified using QUANTI-Star™. This easy-to-use detection reagent contains an ACP5-specific colorimetric substrate, which harbors a phosphate group. Cleavage of this phosphate group by secreted (s)ACP5 leads to the formation of a soluble yellow reaction product, which can be detected at 405 nm using a microplate reader or spectrophotometer. It is therefore combinable with other reporter genes, such as SEAP or Lucia luciferase. ACP5 secretion can be assessed without disturbing cells, thus allowing the repeated sampling of cell cultures for kinetic studies.

For your convenience, InvivoGen offers secreted ACP5 and Lucia-expressing HEK-Lucia-Star™ ARE cells designed to monitor oxidative stress responses. 

Features and advantages of QUANTI-Star™ ACP5 Detection reagent

• Convenient, easy, and rapid
• Cost-effective (1ml = 10 x 96-well plates)
• Short hands-on-time < 10 min
• Easy to prepare: just mix QS reagent and QS buffer with water
• Highly sensitive
• Allows for high-throughput screening (HTS)

Figures

1. In a sterile bottle or flask, prepare QUANTI-Star™ by adding 0.5 ml of QS reagent and 0.5 ml of QS buffer to 49 ml of sterile water. Mix by vortexing and incubate at RT for 10 min before use.

2. Add 80 μl of colorless QUANTI-Star™ solution to 20 μl of the sample (supernatant of ACP5-expressing cells) or negative control (cell culture medium) per well.

3. Incubate at 37°C for 1 h.

3. Add 100 µl of 1M NaOH stop solution (0.1 M final concentration). A color change to yellow will occur. 

4. Measure optical density (OD) at 405 nm using a microplate reader.

Specifications

Cost-effective: 1 ml is sufficient for 10 x 96-well plates (1000 tests)

Practical: After preparation, QUANTI-Star™ is stable for 48 hours at 4°C and 1 month at -20°C.

Quality control:

• Physicochemical characterization (pH, appearance).
• Each lot is thoroughly tested in functional assays.

Contents

QUANTI-Star™ is a two-component reporter kit that contains:

• QUANTI-Star™ reagent (100X)
• QUANTI-Star™ buffer (100X)

QUANTI-Star™ is supplied in two different formats:

• rep-qst1: 0.5 ml of QUANTI-Star™ reagent and 0.5 ml of QUANTI-Star™ buffer, sufficient to prepare QUANTI‑Star™ Solution for 5 x 96-well plates
• rep-qst2: 1 ml of QUANTI-Star™ reagent and 1 ml of QUANTI-Star™ buffer, sufficient to prepare QUANTI‑Star™ Solution for 10 x 96-well plates

 QUANTI-Star™ reagent and  QUANTI-Star™ buffer are shipped at room temperature.

 Upon receipt, store QUANTI-Star™ reagent and  QUANTI-Star™ buffer at -20°C.

 After preparation, the product is stable for 48 hours at 4°C and at least 1 month at -20°C.