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Human Aggrecan (AGC) 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 Aggrecan (AGC) ELISA Kit | |
| Method: | Sandwich | |
| Synonyms: | ACAN; AGC1; AGCAN; CSPG1; CSPCP; CSPGCP; MSK16; SEDK; Cartilage-specific proteoglycan core protein; Large Aggregating Proteoglycan; Chondroitin Sulfate Proteoglycan 1 |
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| Detection range: | 0.468-30ng/mL | |
| Target Protein: | AGC | |
| Size: | 96T/48T | |
| Quality guarantee period: | for 12 months, 16 months | |
| Catalog number: | DL-AGC-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：ACAN; AGC1; AGCAN; CSPG1; CSPCP; CSPGCP; MSK16; SEDK; Cartilage-specific proteoglycan core protein; Large Aggregating Proteoglycan; Chondroitin Sulfate Proteoglycan 1
Function: This proteoglycan is a major component of extracellular matrix of cartilagenous tissues. A major function of this protein is to resist compression in cartilage. It binds avidly to hyaluronic acid via an N-terminal globular region.
Sequence：
10 20 30 40 50 MTTLLWVFVT LRVITAAVTV ETSDHDNSLS VSIPQPSPLR VLLGTSLTIP 60 70 80 90 100 CYFIDPMHPV TTAPSTAPLA PRIKWSRVSK EKEVVLLVAT EGRVRVNSAY 110 120 130 140 150 QDKVSLPNYP AIPSDATLEV QSLRSNDSGV YRCEVMHGIE DSEATLEVVV 160 170 180 190 200 KGIVFHYRAI STRYTLDFDR AQRACLQNSA IIATPEQLQA AYEDGFHQCD 210 220 230 240 250 AGWLADQTVR YPIHTPREGC YGDKDEFPGV RTYGIRDTNE TYDVYCFAEE 260 270 280 290 300 MEGEVFYATS PEKFTFQEAA NECRRLGARL ATTGQLYLAW QAGMDMCSAG 310 320 330 340 350 WLADRSVRYP ISKARPNCGG NLLGVRTVYV HANQTGYPDP SSRYDAICYT 360 370 380 390 400 GEDFVDIPEN FFGVGGEEDI TVQTVTWPDM ELPLPRNITE GEARGSVILT 410 420 430 440 450 VKPIFEVSPS PLEPEEPFTF APEIGATAFA EVENETGEAT RPWGFPTPGL 460 470 480 490 500 GPATAFTSED LVVQVTAVPG QPHLPGGVVF HYRPGPTRYS LTFEEAQQAC 510 520 530 540 550 LRTGAVIASP EQLQAAYEAG YEQCDAGWLR DQTVRYPIVS PRTPCVGDKD 560 570 580 590 600 SSPGVRTYGV RPSTETYDVY CFVDRLEGEV FFATRLEQFT FQEALEFCES 610 620 630 640 650 HNATLATTGQ LYAAWSRGLD KCYAGWLADG SLRYPIVTPR PACGGDKPGV 660 670 680 690 700 RTVYLYPNQT GLPDPLSRHH AFCFRGISAV PSPGEEEGGT PTSPSGVEEW 710 720 730 740 750 IVTQVVPGVA AVPVEEETTA VPSGETTAIL EFTTEPENQT EWEPAYTPVG 760 770 780 790 800 TSPLPGILPT WPPTGAATEE STEGPSATEV PSASEEPSPS EVPFPSEEPS 810 820 830 840 850 PSEEPFPSVR PFPSVELFPS EEPFPSKEPS PSEEPSASEE PYTPSPPVPS 860 870 880 890 900 WTELPSSGEE SGAPDVSGDF TGSGDVSGHL DFSGQLSGDR ASGLPSGDLD 910 920 930 940 950 SSGLTSTVGS GLPVESGLPS GDEERIEWPS TPTVGELPSG AEILEGSASG 960 970 980 990 1000 VGDLSGLPSG EVLETSASGV GDLSGLPSGE VLETTAPGVE DISGLPSGEV 1010 1020 1030 1040 1050 LETTAPGVED ISGLPSGEVL ETTAPGVEDI SGLPSGEVLE TTAPGVEDIS 1060 1070 1080 1090 1100 GLPSGEVLET TAPGVEDISG LPSGEVLETT APGVEDISGL PSGEVLETAA 1110 1120 1130 1140 1150 PGVEDISGLP SGEVLETAAP GVEDISGLPS GEVLETAAPG VEDISGLPSG 1160 1170 1180 1190 1200 EVLETAAPGV EDISGLPSGE VLETAAPGVE DISGLPSGEV LETAAPGVED 1210 1220 1230 1240 1250 ISGLPSGEVL ETAAPGVEDI SGLPSGEVLE TAAPGVEDIS GLPSGEVLET 1260 1270 1280 1290 1300 AAPGVEDISG LPSGEVLETA APGVEDISGL PSGEVLETAA PGVEDISGLP 1310 1320 1330 1340 1350 SGEVLETAAP GVEDISGLPS GEVLETAAPG VEDISGLPSG EVLETAAPGV 1360 1370 1380 1390 1400 EDISGLPSGE VLETAAPGVE DISGLPSGEV LETAAPGVED ISGLPSGEVL 1410 1420 1430 1440 1450 ETTAPGVEEI SGLPSGEVLE TTAPGVDEIS GLPSGEVLET TAPGVEEISG 1460 1470 1480 1490 1500 LPSGEVLETS TSAVGDLSGL PSGGEVLEIS VSGVEDISGL PSGEVVETSA 1510 1520 1530 1540 1550 SGIEDVSELP SGEGLETSAS GVEDLSRLPS GEEVLEISAS GFGDLSGLPS 1560 1570 1580 1590 1600 GGEGLETSAS EVGTDLSGLP SGREGLETSA SGAEDLSGLP SGKEDLVGSA 1610 1620 1630 1640 1650 SGDLDLGKLP SGTLGSGQAP ETSGLPSGFS GEYSGVDLGS GPPSGLPDFS 1660 1670 1680 1690 1700 GLPSGFPTVS LVDSTLVEVV TASTASELEG RGTIGISGAG EISGLPSSEL 1710 1720 1730 1740 1750 DISGRASGLP SGTELSGQAS GSPDVSGEIP GLFGVSGQPS GFPDTSGETS 1760 1770 1780 1790 1800 GVTELSGLSS GQPGISGEAS GVLYGTSQPF GITDLSGETS GVPDLSGQPS 1810 1820 1830 1840 1850 GLPGFSGATS GVPDLVSGTT SGSGESSGIT FVDTSLVEVA PTTFKEEEGL 1860 1870 1880 1890 1900 GSVELSGLPS GEADLSGKSG MVDVSGQFSG TVDSSGFTSQ TPEFSGLPSG 1910 1920 1930 1940 1950 IAEVSGESSR AEIGSSLPSG AYYGSGTPSS FPTVSLVDRT LVESVTQAPT 1960 1970 1980 1990 2000 AQEAGEGPSG ILELSGAHSG APDMSGEHSG FLDLSGLQSG LIEPSGEPPG 2010 2020 2030 2040 2050 TPYFSGDFAS TTNVSGESSV AMGTSGEASG LPEVTLITSE FVEGVTEPTI 2060 2070 2080 2090 2100 SQELGQRPPV THTPQLFESS GKVSTAGDIS GATPVLPGSG VEVSSVPESS 2110 2120 2130 2140 2150 SETSAYPEAG FGASAAPEAS REDSGSPDLS ETTSAFHEAN LERSSGLGVS 2160 2170 2180 2190 2200 GSTLTFQEGE ASAAPEVSGE STTTSDVGTE APGLPSATPT ASGDRTEISG 2210 2220 2230 2240 2250 DLSGHTSQLG VVISTSIPES EWTQQTQRPA ETHLEIESSS LLYSGEETHT 2260 2270 2280 2290 2300 VETATSPTDA SIPASPEWKR ESESTAAAPA RSCAEEPCGA GTCKETEGHV 2310 2320 2330 2340 2350 ICLCPPGYTG EHCNIDQEVC EEGWNKYQGH CYRHFPDRET WVDAERRCRE 2360 2370 2380 2390 2400 QQSHLSSIVT PEEQEFVNNN AQDYQWIGLN DRTIEGDFRW SDGHPMQFEN 2410 2420 2430 2440 2450 WRPNQPDNFF AAGEDCVVMI WHEKGEWNDV PCNYHLPFTC KKGTVACGEP 2460 2470 2480 2490 2500 PVVEHARTFG QKKDRYEINS LVRYQCTEGF VQRHMPTIRC QPSGHWEEPQ 2510 2520 2530 ITCTDPTTYK RRLQKRSSRH PRRSRPSTAH
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of AGC in human serum, plasma or other biological fluids.
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 AGC is typically less than 0.16ng/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 AGC.
No significant cross-reactivity or interference between AGC and analogues was observed.
You can reference link of the kit as following
Introduction
| Item | Standard | Test | |
| Description |
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of AGC in human serum, plasma 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 AGC 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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