Human Acid Phosphatase 5, Tartrate Resistant (ACP5) (Sandwich ELISA) ELISA Kit

Mfr.No.

SEA902Hu

Sensitivity

0.129ng/mL

Detection Range

0.312-20ng/mL

Reactivity

Human

Assay Duration

Assay Type

Sandwich

Description

This assay has high sensitivity and excellent specificity for detection of Acid Phosphatase 5, Tartrate Resistant (ACP5).
No significant cross-reactivity or interference between Acid Phosphatase 5, Tartrate Resistant (ACP5) and analogues was observed.

Alternative Names

TRAP; TrATPase; Tartrate Resistant Acid Phosphatase; Type 5 acid phosphatase

Detection Method

Colormetric

Kit Components

Reagents	Quantity	Reagents	Quantity
Pre-coated, ready to use 96-well strip plate	1	Plate sealer for 96 wells	4
Standard	2	Standard Diluent	1×20mL
Detection Reagent A	1×120µL	Assay Diluent A	1×12mL
Detection Reagent B	1×120µL	Assay Diluent B	1×12mL
TMB Substrate	1×9mL	Stop Solution	1×6mL
Wash Buffer (30 × concentrate)	1×20mL	Instruction manual	1

Recovery

Matrices listed below were spiked with certain level of recombinant Acid Phosphatase 5, Tartrate Resistant (ACP5) and the recovery rates were calculated by comparing the measured value to the expected amount of Acid Phosphatase 5, Tartrate Resistant (ACP5) in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	97-104	101
EDTA plasma(n=5)	89-96	92
heparin plasma(n=5)	78-95	83

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Acid Phosphatase 5, Tartrate Resistant (ACP5) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1:2	1:4	1:8	1:16
serum(n=5)	91-98%	80-92%	78-104%	89-103%
EDTA plasma(n=5)	99-105%	90-99%	88-102%	89-101%
heparin plasma(n=5)	85-101%	81-101%	84-101%	97-105%

Features

Detects protein (regular version)

Species

Human

Precautions

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Acid Phosphatase 5, Tartrate Resistant (ACP5) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Acid Phosphatase 5, Tartrate Resistant (ACP5) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Notes

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Acid Phosphatase 5, Tartrate Resistant (ACP5). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Acid Phosphatase 5, Tartrate Resistant (ACP5). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Acid Phosphatase 5, Tartrate Resistant (ACP5), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Acid Phosphatase 5, Tartrate Resistant (ACP5) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Usage Notes

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

Protocol

Datasheet

Gene ID	54
UniProt ID	P13686

Applications	ELISA
Research Area	Enzyme///Kinase

Storage	4°C for 1 month/ -20°C for long-term(within shelf life)
Shelf Life	one year

Reference	Valvular osteoclasts in calcification and aortic valve stenosis severity.

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ELISA Kit for Acid Phosphatase 5, Tartrate Resistant (ACP5) (Human)

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