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Human Von Willebrand Factor A Domain Containing Protein 3A (vWA3A) 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 Von Willebrand Factor A Domain Containing Protein 3A (vWA3A) ELISA Kit | |
| Method: | Sandwich | |
| Synonyms: | ||
| Detection range: | 0.468-30ng/mL | |
| Target Protein: | vWA3A | |
| Size: | 96T/48T | |
| Quality guarantee period: | for 12 months, 16 months | |
| Catalog number: | DL-vWA3A-Hu (traditional) | (ready-to-use) |
| Assay length | 1-4.5Hours | 1-3.5Hours |
| Advantages: |
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| Instruction Manual |
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1.Overview：
Sequence：
10 20 30 40 50 MKKYRKISIG CFAMATQTSH VFHGQENMFL ENHCIRRNTG RDSKKPLKQK 60 70 80 90 100 NMNGLGQNSD NGLLVTHVNQ TQDLLRLQGS ETQSSDWEDS EDWLSAHSLK 110 120 130 140 150 CQKLTLADLI SQGTEVLEEG TNVVQKICFS TQIIRHFESK LSDTIEVYQE 160 170 180 190 200 RIQWLTENSK KAFGLIKGAR VSILIDVSAI SSGPQKEEFQ KDLMSLIDEQ 210 220 230 240 250 LSHKEKLFVL SFGTNAGSLW PDPMEVSAST LQELKLWVKT LQPDGGSNLL 260 270 280 290 300 QALKKIFTLK GLDSLVAIMR SCPDQPSEIL SDYIQQSTMG RDLIIHFITY 310 320 330 340 350 RCDDQMPPAV LKNLAEAVRG YYHCYSPKME HYTSRDMDEL LAEIQKAQSL 360 370 380 390 400 LSHVQALQHS SPCEALTCTM EEISTEITNG PLISLLPKPP KHDAPLTIEF 410 420 430 440 450 PNLDKTSAEW LKVNGLKAKK LSLYQVLAPN AFSPVEEFVP ILQKTVSSTI 460 470 480 490 500 HEKAMIQFEW HDGTVKNIHV DPPFLYKYQQ QLSRAMRMYE RRIEWLSLAS 510 520 530 540 550 RRIWGTVCEK RVVVLLDISA TNSMYIIHIQ HSLRLLLEEQ LSNKDCFNLI 560 570 580 590 600 AFGSTIESWR PEMVPVSHNN LQSAWRWALN LRCRGSRNVL SALRKAVEVD 610 620 630 640 650 FKDKDKHQSQ GIYLFTGGIP DQDMPTLSAY MAEACGGCDL QLNVCLFYVG 660 670 680 690 700 EPKMDTTPPA RYASHTDTAA AYKEVTRAAG GRFHWFGDTG IYESDDINSI 710 720 730 740 750 MSEMEKALNY SQKCAFLMAS LKNHSGKVLG SSALPKEKPK TLQLRSQPKK 760 770 780 790 800 LCPPRPTVPL GARMSIKDDP DREKSPPLKS LKWRPLSSRV GISPAAAQPT 810 820 830 840 850 KEGMMELRRK TKSREAETSL LLFYTEKGND VGSVYKKYPQ GRGLRRTSSS 860 870 880 890 900 IDLPRKDTVC SSQEWVAKYG LKKLKLEISR CMGPNCTHQK SGQRSASAKH 910 920 930 940 950 CSIFPSVEIH GVVRHIQWTP REMEVYIRHL EKVLRRYVQR LQWLLSGSRR 960 970 980 990 1000 LFGTVLESKV CILLDTSGSM GPYLQQVKTE LVLLIWEQLR KCCDSFNLLS 1010 1020 1030 1040 1050 FAESFQSWQD TLVETTDAAC HEAMQWVTHL QAQGSTSILQ ALLKAFSFHD 1060 1070 1080 1090 1100 LEGLYLLTDG KPDTSCSLVL NEVQKLREKR DVKVHTISLN CSDRAAVEFL 1110 1120 1130 1140 1150 RKLASFTGGR YHCPVGEDTL SKIHSLLTKG FINEKDPTLP PFEGDDLRIL 1160 1170 1180 AQEITKARSF LWQAQSFRSQ LQKKNDAEPK VTLS
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of vWA3A in human plasma.
DETECTION RANGE
0.468-30ng/mL. The standard curve concentrations used for the ELISA’s were 30ng/mL, 15ng/mL, 7.5ng/mL, 3.75ng/mL, 1.87ng/mL, 0.937ng/mL, 0.468ng/mL.
SENSITIVITY
The minimum detectable dose of vWA3A is typically less than 0.15ng/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 vWA3A.
No significant cross-reactivity or interference between vWA3A and analogues was observed.
You can reference link of the kit as following
https://dldevelop.com/Research-reagent/dl-vwa3a-hu.html
https://www.dldevelop.com/uploadfile/data/DL-vWA3A-Hu.pdf
Introduction
| Item | Standard | Test | |
| Description |
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of vWA3A in human plasma. |
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 vWA3A 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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