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Human UDP-Glucose Glycoprotein Glucosyltransferase 1 (UGGT1) 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 UDP-Glucose Glycoprotein Glucosyltransferase 1 (UGGT1) ELISA Kit | |
| Method: | Sandwich | |
| Synonyms: | UGTR; HUGT1; GT; UGCGL1; UGGT; UGT1; UDP-glucose ceramide glucosyltransferase-like 1 |
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| Detection range: | 0.312-20ng/mL | |
| Target Protein: | ||
| Size: | 96T/48T | |
| Quality guarantee period: | for 12 months, 16 months | |
| Catalog number: | DL-UGGT1-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：UGTR; HUGT1; GT; UGCGL1; UGGT; UGT1; UDP-glucose ceramide glucosyltransferase-like 1
Function: Recognizes glycoproteins with minor folding defects. Reglucosylates single N-glycans near the misfolded part of the protein, thus providing quality control for protein folding in the endoplasmic reticulum. Reglucosylated proteins are recognized by calreticulin for recycling to the endoplasmic reticulum and refolding or degradation.
Sequence：
10 20 30 40 50 MGCKGDASGA CAAGALPVTG VCYKMGVLVV LTVLWLFSSV KADSKAITTS 60 70 80 90 100 LTTKWFSTPL LLEASEFLAE DSQEKFWNFV EASQNIGSSD HDGTDYSYYH 110 120 130 140 150 AILEAAFQFL SPLQQNLFKF CLSLRSYSAT IQAFQQIAAD EPPPEGCNSF 160 170 180 190 200 FSVHGKKTCE SDTLEALLLT ASERPKPLLF KGDHRYPSSN PESPVVIFYS 210 220 230 240 250 EIGSEEFSNF HRQLISKSNA GKINYVFRHY IFNPRKEPVY LSGYGVELAI 260 270 280 290 300 KSTEYKAKDD TQVKGTEVNT TVIGENDPID EVQGFLFGKL RDLHPDLEGQ 310 320 330 340 350 LKELRKHLVE STNEMAPLKV WQLQDLSFQT AARILASPVE LALVVMKDLS 360 370 380 390 400 QNFPTKARAI TKTAVSSELR TEVEENQKYF KGTLGLQPGD SALFINGLHM 410 420 430 440 450 DLDTQDIFSL FDVLRNEARV MEGLHRLGIE GLSLHNVLKL NIQPSEADYA 460 470 480 490 500 VDIRSPAISW VNNLEVDSRY NSWPSSLQEL LRPTFPGVIR QIRKNLHNMV 510 520 530 540 550 FIVDPAHETT AELMNTAEMF LSNHIPLRIG FIFVVNDSED VDGMQDAGVA 560 570 580 590 600 VLRAYNYVAQ EVDDYHAFQT LTHIYNKVRT GEKVKVEHVV SVLEKKYPYV 610 620 630 640 650 EVNSILGIDS AYDRNRKEAR GYYEQTGVGP LPVVLFNGMP FEREQLDPDE 660 670 680 690 700 LETITMHKIL ETTTFFQRAV YLGELPHDQD VVEYIMNQPN VVPRINSRIL 710 720 730 740 750 TAERDYLDLT ASNNFFVDDY ARFTILDSQG KTAAVANSMN YLTKKGMSSK 760 770 780 790 800 EIYDDSFIRP VTFWIVGDFD SPSGRQLLYD AIKHQKSSNN VRISMINNPA 810 820 830 840 850 KEISYENTQI SRAIWAALQT QTSNAAKNFI TKMAKEGAAE ALAAGADIAE 860 870 880 890 900 FSVGGMDFSL FKEVFESSKM DFILSHAVYC RDVLKLKKGQ RAVISNGRII 910 920 930 940 950 GPLEDSELFN QDDFHLLENI ILKTSGQKIK SHIQQLRVEE DVASDLVMKV 960 970 980 990 1000 DALLSAQPKG DPRIEYQFFE DRHSAIKLRP KEGETYFDVV AVVDPVTREA 1010 1020 1030 1040 1050 QRLAPLLLVL AQLINMNLRV FMNCQSKLSD MPLKSFYRYV LEPEISFTSD 1060 1070 1080 1090 1100 NSFAKGPIAK FLDMPQSPLF TLNLNTPESW MVESVRTPYD LDNIYLEEVD 1110 1120 1130 1140 1150 SVVAAEYELE YLLLEGHCYD ITTGQPPRGL QFTLGTSANP VIVDTIVMAN 1160 1170 1180 1190 1200 LGYFQLKANP GAWILRLRKG RSEDIYRIYS HDGTDSPPDA DEVVIVLNNF 1210 1220 1230 1240 1250 KSKIIKVKVQ KKADMVNEDL LSDGTSENES GFWDSFKWGF TGQKTEEVKQ 1260 1270 1280 1290 1300 DKDDIINIFS VASGHLYERF LRIMMLSVLK NTKTPVKFWF LKNYLSPTFK 1310 1320 1330 1340 1350 EFIPYMANEY NFQYELVQYK WPRWLHQQTE KQRIIWGYKI LFLDVLFPLV 1360 1370 1380 1390 1400 VDKFLFVDAD QIVRTDLKEL RDFNLDGAPY GYTPFCDSRR EMDGYRFWKS 1410 1420 1430 1440 1450 GYWASHLAGR KYHISALYVV DLKKFRKIAA GDRLRGQYQG LSQDPNSLSN 1460 1470 1480 1490 1500 LDQDLPNNMI HQVPIKSLPQ EWLWCETWCD DASKKRAKTI DLCNNPMTKE 1510 1520 1530 1540 1550 PKLEAAVRIV PEWQDYDQEI KQLQIRFQKE KETGALYKEK TKEPSREGPQ KREEL
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of UGGT1 in human tissue homogenates, cell lysates or 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 UGGT1 is typically less than 0.113ng/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 UGGT1.
No significant cross-reactivity or interference between UGGT1 and analogues was observed.
You can reference link of the kit as following
https://dldevelop.com/Research-reagent/dl-uggt1-hu.html
https://www.dldevelop.com/uploadfile/data/DL-UGGT1-Hu.pdf
Introduction
| Item | Standard | Test | |
| Description |
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of UGGT1 in human tissue homogenates, cell lysates 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 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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