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Human ATPase, Cu++ Transporting Beta Polypeptide (ATP7b) ELISA Kit
two product lines: Traditional ELISA Kit and Ready-to-Use ELISA Kit.
| Traditional ELISA Kit | Ready-to-Use ELISA KIT | |
| Product name: | Human ATPase, Cu++ Transporting Beta Polypeptide (ATP7b) ELISA Kit | |
| Method: | Sandwich | |
| Synonyms: | PWD; WC1; WD; WND; Wilson Disease Protein; Copper pump 2; Wilson disease-associated protein |
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| Detection range: | 0.156-10ng/mL | |
| Target Protein: | ATP7b | |
| Size: | 96T/48T | |
| Quality guarantee period: | for 12 months, 16 months | |
| Catalog number: | DL-ATP7b-Hu (traditional) | (ready-to-use) |
| Assay length | 1-4.5Hours | 1-3.5Hours |
| Advantages: |
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| Instruction Manual |
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Other names：PWD; WC1; WD; WND; Wilson Disease Protein; Copper pump 2; Wilson disease-associated protein
Function: Involved in the export of copper out of the cells, such as the efflux of hepatic copper into the bile.
Sequence：
10 20 30 40 50 MPEQERQITA REGASRKILS KLSLPTRAWE PAMKKSFAFD NVGYEGGLDG 60 70 80 90 100 LGPSSQVATS TVRILGMTCQ SCVKSIEDRI SNLKGIISMK VSLEQGSATV 110 120 130 140 150 KYVPSVVCLQ QVCHQIGDMG FEASIAEGKA ASWPSRSLPA QEAVVKLRVE 160 170 180 190 200 GMTCQSCVSS IEGKVRKLQG VVRVKVSLSN QEAVITYQPY LIQPEDLRDH 210 220 230 240 250 VNDMGFEAAI KSKVAPLSLG PIDIERLQST NPKRPLSSAN QNFNNSETLG 260 270 280 290 300 HQGSHVVTLQ LRIDGMHCKS CVLNIEENIG QLLGVQSIQV SLENKTAQVK 310 320 330 340 350 YDPSCTSPVA LQRAIEALPP GNFKVSLPDG AEGSGTDHRS SSSHSPGSPP 360 370 380 390 400 RNQVQGTCST TLIAIAGMTC ASCVHSIEGM ISQLEGVQQI SVSLAEGTAT 410 420 430 440 450 VLYNPSVISP EELRAAIEDM GFEASVVSES CSTNPLGNHS AGNSMVQTTD 460 470 480 490 500 GTPTSVQEVA PHTGRLPANH APDILAKSPQ STRAVAPQKC FLQIKGMTCA 510 520 530 540 550 SCVSNIERNL QKEAGVLSVL VALMAGKAEI KYDPEVIQPL EIAQFIQDLG 560 570 580 590 600 FEAAVMEDYA GSDGNIELTI TGMTCASCVH NIESKLTRTN GITYASVALA 610 620 630 640 650 TSKALVKFDP EIIGPRDIIK IIEEIGFHAS LAQRNPNAHH LDHKMEIKQW 660 670 680 690 700 KKSFLCSLVF GIPVMALMIY MLIPSNEPHQ SMVLDHNIIP GLSILNLIFF 710 720 730 740 750 ILCTFVQLLG GWYFYVQAYK SLRHRSANMD VLIVLATSIA YVYSLVILVV 760 770 780 790 800 AVAEKAERSP VTFFDTPPML FVFIALGRWL EHLAKSKTSE ALAKLMSLQA 810 820 830 840 850 TEATVVTLGE DNLIIREEQV PMELVQRGDI VKVVPGGKFP VDGKVLEGNT 860 870 880 890 900 MADESLITGE AMPVTKKPGS TVIAGSINAH GSVLIKATHV GNDTTLAQIV 910 920 930 940 950 KLVEEAQMSK APIQQLADRF SGYFVPFIII MSTLTLVVWI VIGFIDFGVV 960 970 980 990 1000 QRYFPNPNKH ISQTEVIIRF AFQTSITVLC IACPCSLGLA TPTAVMVGTG 1010 1020 1030 1040 1050 VAAQNGILIK GGKPLEMAHK IKTVMFDKTG TITHGVPRVM RVLLLGDVAT 1060 1070 1080 1090 1100 LPLRKVLAVV GTAEASSEHP LGVAVTKYCK EELGTETLGY CTDFQAVPGC 1110 1120 1130 1140 1150 GIGCKVSNVE GILAHSERPL SAPASHLNEA GSLPAEKDAV PQTFSVLIGN 1160 1170 1180 1190 1200 REWLRRNGLT ISSDVSDAMT DHEMKGQTAI LVAIDGVLCG MIAIADAVKQ 1210 1220 1230 1240 1250 EAALAVHTLQ SMGVDVVLIT GDNRKTARAI ATQVGINKVF AEVLPSHKVA 1260 1270 1280 1290 1300 KVQELQNKGK KVAMVGDGVN DSPALAQADM GVAIGTGTDV AIEAADVVLI 1310 1320 1330 1340 1350 RNDLLDVVAS IHLSKRTVRR IRINLVLALI YNLVGIPIAA GVFMPIGIVL 1360 1370 1380 1390 1400 QPWMGSAAMA ASSVSVVLSS LQLKCYKKPD LERYEAQAHG HMKPLTASQV 1410 1420 1430 1440 1450 SVHIGMDDRW RDSPRATPWD QVSYVSQVSL SSLTSDKPSR HSAAADDDGD 1460 KWSLLLNGRD EEQYI
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of ATP7b in human tissue homogenates and other biological fluids.
DETECTION RANGE
0.156-10ng/mL. The standard curve concentrations used for the ELISA’s were 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.312ng/mL, 0.156ng/mL.
SENSITIVITY
The minimum detectable dose of ATP7b is typically less than 0.056ng/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 ATP7b.
No significant cross-reactivity or interference between ATP7b and analogues was observed.
You can reference link of the kit as following
https://dldevelop.com/Research-reagent/dl-atp7b-hu.html
https://dldevelop.com/Research-reagent/dl-atp7b-mu.html
Introduction
| Item | Standard | Test | |
| Description |
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of ATP7b in human serum, plasma,tissue homogenates and other biological fluids. |
Conform | |
| Identification | Colorimetric | Positive | |
| Composition | Traditional ELISA Kit | Ready-to-Use ELISA KIT | Conform |
| Pre-coated, ready to use 96-well strip plate 1 | Pre-coated, ready to use 96-well strip plate 1 | ||
| Plate sealer for 96 wells 2 | Plate sealer for 96 wells 2 | ||
| Standard 2 | Standard 2 | ||
| Diluents buffer 1×45mL | Standard Diluent 1×20mL | ||
| Detection Reagent A 1×120μL | Detection Solution A 1×12mL | ||
| Detection Reagent B 1×120μL | Detection Solution B 1×12mL | ||
| TMB Substrate 1×9mL | TMB Substrate 1×9mL | ||
| Stop Solution 1×6mL | Stop Solution 1×6mL | ||
| Wash Buffer (30 × concentrate) 1×20mL | Wash Buffer (30 × concentrate) 1×20mL | ||
| Instruction manual 1 | Instruction manual 1 | ||
Test principle
The microtiter plate provided in this kit has been pre-coated with an antibody specific to the index. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to the index. 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 the index, 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 the index in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Recovery
Matrices listed below were spiked with certain level of recombinant ATP7b and the recovery rates were calculated by comparing the measured value to the expected amount of the index in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 81-93 | 86 |
| EDTA plasma(n=5) | 80-97 | 88 |
| heparin plasma(n=5) | 90-101 | 95 |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of the index 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) | 82-96% | 83-98% | 81-99% | 93-101% |
| EDTA plasma(n=5) | 88-101% | 86-95% | 90-102% | 80-93% |
| heparin plasma(n=5) | 80-91% | 82-90% | 95-104% | 79-95% |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level the index were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level the index 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 37℃;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37℃;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 1 hour at 37℃;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37℃;
8. Add 50µL Stop Solution. Read at 450nm immediately.
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