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Mouse Bromodomain-Containing Protein 4 (BRD4) ELISA Kit
two product lines: Traditional ELISA Kit and Ready-to-Use ELISA Kit.
| Traditional ELISA Kit | Ready-to-Use ELISA KIT | |
| Product name: | Mouse Bromodomain-Containing Protein 4 (BRD4) ELISA Kit | |
| Method: | Sandwich | |
| Synonyms: | Mitotic chromosome-associated protein(MCAP) |
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| Detection range: | 31.2-2000pg/ml | |
| Target Protein: | BRD4 | |
| Size: | 96T/48T | |
| Quality guarantee period: | for 12 months, 16 months | |
| Catalog number: | DL-BRD4-Mu (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：Mitotic chromosome-associated protein(MCAP)
Function: Plays a role in a process governing chromosomal dynamics during mitosis.
Sequence：
10 20 30 40 50 MSTESGPGTR LRNLPVMGDG LETSQMSTTQ AQAQPQPANA ASTNPPPPET 60 70 80 90 100 SNPNKPKRQT NQLQYLLRVV LKTLWKHQFA WPFQQPVDAV KLNLPDYYKI 110 120 130 140 150 IKTPMDMGTI KKRLENNYYW NAQECIQDFN TMFTNCYIYN KPGDDIVLMA 160 170 180 190 200 EALEKLFLQK INELPTEETE IMIVQAKGRG RGRKETGAAK PGVSTVPNTT 210 220 230 240 250 QASTSPQTQT PQQNPPPPVQ ATTHPFPAVT PDLIAQPPVM TMVPPQPLQT 260 270 280 290 300 PSPVPPQPPP PPAPVPQPVQ SHPPIIATTP QPVKTKKGVK RKADTTTPTT 310 320 330 340 350 IDPIHEPPSL APEPKTAKLG PRRESSRPVK PPKKDVPDSQ QHPGPEKSSK 360 370 380 390 400 ISEQLKCCSG ILKEMFAKKH AAYAWPFYKP VDVEALGLHD YCDIIKHPMD 410 420 430 440 450 MSTIKSKLES REYRDAQEFG ADVRLMFSNC YKYNPPDHEV VAMARKLQDV 460 470 480 490 500 FEMRFAKMPD EPEEPVVTVS SPAVPPPTKV VAPPSSSDSS SDSSSDSDSS 510 520 530 540 550 TDDSEEERAQ RLAELQEQLK AVHEQLAALS QPQQNKPKKK EKDKKEKKKE 560 570 580 590 600 KHKKKEEVEE NKKSKTKELP PKKTKKNNSS NSNVSKKEPV PTKTKPPPTY 610 620 630 640 650 ESEEEDKCKP MSYEEKRQLS LDINKLPGEK LGRVVHIIQS REPSLKNSNP 660 670 680 690 700 DEIEIDFETL KPSTLRELER YVTSCLRKKR KPQAEKVDVI AGSSKMKGFS 710 720 730 740 750 SSESESTSES SSSDSEDSET EMAPKSKKKG HTGRDQKKHH HHHHPQMQPA 760 770 780 790 800 PAPVPQQPPP PPQQPPPPPP PQQQQQQPPP PPPPPSMPQQ TAPAMKSSPP 810 820 830 840 850 PFITAQVPVL EPQLPGSVFD PIGHFTQPIL HLPQPELPPH LPQPPEHSTP 860 870 880 890 900 PHLNQHAVVS PPALHNALPQ QPSRPSNRAA ALPPKPTRPP AVSPALAQPP 910 920 930 940 950 LLPQPPMAQP PQVLLEDEEP PAPPLTSMQM QLYLQQLQKV QPPTPLLPSV 960 970 980 990 1000 KVQSQPPPPL PPPPHPSVQQ QQLQPQPPPP PPPQPQPPPQ QQHQPPPRPV 1010 1020 1030 1040 1050 HLPSMPFSAH IQQPPPPPGQ QPTHPPPGQQ PPPPQPAKPQ QVIQHHPSPR 1060 1070 1080 1090 1100 HHKSDPYSAG HLREAPSPLM IHSPQMPQFQ SLTHQSPPQQ NVQPKKQVKG 1110 1120 1130 1140 1150 RAEPQPPGPV MGQGQGCPPA SPAAVPMLSQ ELRPPSVVQP QPLVVVKEEK 1160 1170 1180 1190 1200 IHSPIIRSEP FSTSLRPEPP KHPENIKAPV HLPQRPEMKP VDIGRPVIRP 1210 1220 1230 1240 1250 PEQSAPPPGA PDKDKQKQEP KTPVAPKKDL KIKNMGSWAS LVQKHPTTPS 1260 1270 1280 1290 1300 STAKSSSDSF EHFRRAAREK EEREKALKAQ AEHAEKEKER LRQERMRSRE 1310 1320 1330 1340 1350 DEDALEQARR AHEEARRRQE QQQQQQQQRQ EQQQQQQQAA AVAAASAPQA 1360 1370 1380 1390 1400 QSSQPQSMLD QQRELARKRE QERRRREAMA ATIDMNFQSD LLSIFEENLF
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of BRD4 in mouse serum, plasma, tissue homogenates or other biological fluids.
DETECTION RANGE
31.2-2000pg/ml. The standard curve concentrations used for the ELISA’s were 2000pg/ml, 1000pg/ml, 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml, 31.2pg/ml.
SENSITIVITY
The minimum detectable dose of BRD4 is typically less than 12.5pg/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 BRD4.
No significant cross-reactivity or interference between BRD4 and analogues was observed.
You can reference link of the kit as following
https://dldevelop.com/Research-reagent/dl-brd4-mu.html
https://www.dldevelop.com/uploadfile/data/DL-BRD4-Mu.pdf
Introduction
| Item | Standard | Test | |
| Description |
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of BRD4 in mouse serum, plasma, tissue homogenates or 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 BRD4 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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