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

DL-ACP5-Hu

1. Overview

Other names:TRAP; TrATPase; Tartrate Resistant Acid Phosphatase; Type 5 acid phosphatase

Function: Involved in osteopontin/bone sialoprotein dephosphorylation. Its expression seems to increase in certain pathological states such as Gaucher and Hodgkin diseases, the hairy cell, the B-cell, and the T-cell leukemias.

Sequence

MDMWTALLIL  QALLLPSLAD  GATPALRFVA  VGDWGGVPNA  PFHTAREMAN

AKEIARTVQI  LGADFILSLG  DNFYFTGVQD  INDKRFQETF  EDVFSDRSLR

KVPWYVLAGN  HDHLGNVSAQ  IAYSKISKRW  NFPSPFYRLH  FKIPQTNVSV

AIFMLDTVTL  CGNSDDFLSQ  QPERPRDVKL  ARTQLSWLKK  QLAAAREDYV

LVAGHYPVWS  IAEHGPTHCL  VKQLRPLLAT  YGVTAYLCGH  DHNLQYLQDE

NGVGYVLSGA  GNFMDPSKRH  QRKVPNGYLR  FHYGTEDSLG  GFAYVEISSK

EMTVTYIEAS  GKSLFKTRLP  RRARP

2. Features

INTENDED USE

The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of ACP5 in human serum, plasma, tissue homogenates and other biological fluids.

DETECTION RANGE

0.312-20ng/mL. The standard curve concentrations used for the ELISA’s were 20ng/mL, 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.312ng/mL.

SENSITIVITY

The minimum detectable dose of ACP5 is typically less than 0.121ng/mL.

The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.

SPECIFICITY

This assay has high sensitivity and excellent specificity for detection of ACP5.

No significant cross-reactivity or interference between ACP5 and analogues was observed.

Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between ACP5 and all analogues, therefore, cross reactivity may still exist.

PRECISION

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level ACP5 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level ACP5 were tested on 3 different plates, 8 replicates in each plate.

CV(%) = SD/meanX100

Intra-Assay: CV<10%

Inter-Assay: CV<12%

STABILITY

The stability of ELISA 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 conditions.

Note: To minimize unnecessary influences on the performance, operation procedures and lab conditions, especially room temperature, air humidity, and incubator temperatures should be strictly regulated. It is also strongly suggested that the whole assay is performed by the same experimenter from the beginning to the end.

ASSAY PROCEDURE SUMMARY

1.Prepare all reagents, samples and standards;
2.Add 100µL standard or sample to each well. Incubate 2 hours at 37oC;
3.Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37oC;
4.Aspirate and wash 3 times;
5.Add 100µL prepared Detection Reagent B. Incubate 1 hour at 37oC;
6.Aspirate and wash 5 times;
7.Add 90µL Substrate Solution. Incubate 15-25 minutes at 37oC;
8.Add 50µL Stop Solution. Read at 450nm immediately.

IMPORTANT NOTES

1.Limited by the current conditions and scientific technology, it is impossible to conduct comprehensive identification and analysis tests on the raw materials provided by suppliers. As a result, it is possible there are some qualitative and/or technical risks.
2.The final experimental results will be closely related to the validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available to obtain accurate results.
3.Kits from different batches may be a little different in detection range, sensitivity and color developing time. Please perform the experiment exactly according to the instruction manual included in your kit. Electronic ones on our website are for reference only.
4.Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
5.Protect all reagents from strong light during storage and incubation. All bottle caps of reagents should be closed tightly to prevent evaporation of liquids and contamination by microorganisms.
6.There may be a foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
7.Incorrect procedures during reagent preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
8.Even the same experimenter may get different results from two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before the general assay for each batch is recommended.
9.Each kit has undergone several rigorous quality control tests. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipment. Intra-assay variance among kits from different batches could arise from the above factors as well.
10.Kits from different manufacturers with the same item might produce different results, since we have not compared our products with other manufacturers.
11.The standard of the kit and immunogen used for antibody preparation are commonly recombinant proteins. Different expressed sequence, expression systems, purification methods might be used in recombinant protein preparation. Besides, there might exist differences on the screening technique of antibody and antibody pairs in our kit. Thus we can not guarantee the kit could detect recombinant protein from other companies. So, it is not recommended to use the kit for the detection of recombinant protein.
12.Validity period: 12 months.
13.The instruction manual also works with the 48T kit, but all reagents in the 48T kit are reduced by half.

PRECAUTION

The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this reagent.

You can reference link of the kit as following

http://www.dldevelop.com/Research-reagent/dl-acp5-hu.html

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Post time: Apr-13-2018
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