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Mouse B-Cell CLL/Lymphoma 9 (Bcl9) 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 B-Cell CLL/Lymphoma 9 (Bcl9) ELISA Kit | |
| Method: | Sandwich | |
| Synonyms: | LGS; Protein legless homolog |
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| Detection range: | 78.125-5000pg/mL | |
| Target Protein: | Bcl9 | |
| Size: | 96T/48T | |
| Quality guarantee period: | for 12 months, 16 months | |
| Catalog number: | DL-Bcl9-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：LGS; Protein legless homolog
Function: Involved in signal transduction through the Wnt pathway. Promotes beta-catenin's transcriptional activity.
Sequence：
MHSSNPKVRS SPSGNTQSSP KSKQEVMVRP PTVMSPSGNP QLDSKFSNQG
KQGGSASQSQ PSPCDSKSGG HTPKALPGPG GSMGLKNGAG NGAKGKGKRE
RSISADSFDQ RDPGTPNDDS DIKECNSADH IKSQDSQHTP HSMTPSNATA
PRSSTPSHGQ TTATEPTPAQ KTPAKVVYVF STEMANKAAE AVLKGQVETI
VSFHIQNISN NKTERSTAPL NTQISALRND PKPLPQQPPA PANQDQNSSQ
NTRLQPTPPI PAPAPKPAAP PRPLDRESPG VENKLIPSVG SPASSTPLPP
DGTGPNSTPN NRAVTPVSQG SNSSSADPKA PPPPPVSSGE PPTLGENPDG
LSQEQLEHRE RSLQTLRDIQ RMLFPDEKEF TGAQSGGPQQ NPGVLDGPQK
KPEGPIQAMM AQSQSLGKGP GPRTDVGAPF GPQGHRDVPF SPDEMVPPSM
NSQSGTIGPD HLDHMTPEQI AWLKLQQEFY EEKRRKQEQV VVQQCSLQDM
MVHQHGPRGV VRGPPPPYQM TPSEGWAPGG TEPFSDGINM PHSLPPRGMA
PHPNMPGSQM RLPGFAGMIN SEMEGPNVPN PASRPGLSGV SWPDDVPKIP
DGRNFPPGQG IFSGPGRGER FPNPQGLSEE MFQQQLAEKQ LGLPPGMAME
GIRPSMEMNR MIPGSQRHME PGNNPIFPRI PVEGPLSPSR GDFPKGIPPQ
MGPGRELEFG MVPSGMKGDV NLNVNMGSNS QMIPQKMREA GAGPEEMLKL
RPGGSDMLPA QQKMVPLPFG EHPQQEYGMG PRPFLPMSQG PGSNSGLRNL
REPIGPDQRT NSRLSHMPPL PLNPSSNPTS LNTAPPVQRG LGRKPLDISV
AGSQVHSPGI NPLKSPTMHQ VQSPMLGSPS GNLKSPQTPS QLAGMLAGPA
AAASIKSPPV LGSAAASPVH LKSPSLPAPS PGWTSSPKPP LQSPGIPPNH
KAPLTMASPA MLGNVESGGP PPPTASQPAS VNIPGSLPSS TPYTMPPEPT
LSQNPLSIMM SRMSKFAMPS STPLYHDAIK TVASSDDDSP PARSPNLPSM
NNMPGMGINT QNPRISGPNP VVPMPTLSPM GMTQPLSHSN QMPSPNAVGP
NIPPHGVPMG PGLMSHNPIM GHGSQEPPMV PQGRMGFPQG FPPVQSPPQQ
VPFPHNGPSG GQGSFPGGMG FPGEGPLGRP SNLPQSSADA ALCKPGGPGG
PDSFTVLGNS MPSVFTDPDL QEVIRPGATG IPEFDLSRII PSEKPSQTLQ
YFPRGEVPGR KQPQGPGPGF SHMQGMMGEQ APRMGLALPG MGGPGPVGTP
DIPLGTAPSM PGHNPMRPPA FLQQGMMGPH HRMMSPAQST MPGQPTLMSN
PAAAVGMIPG KDRGPAGLYT HPGPVGSPGM MMSMQGMMGP QQNIMIPPQM
RPRGMAADVG MGGFSQGPGN PGNMMF
2. Features
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of Bcl9 in mouse serum, plasma, tissue homogenates and other biological fluids.
DETECTION RANGE
78.125-5000pg/mL. The standard curve concentrations used for the ELISA’s were 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312pg/mL, 156pg/mL, 78.1pg/mL.
SENSITIVITY
The minimum detectable dose of Bcl9 is typically less than 27pg/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 Bcl9.
No significant cross-reactivity or interference between Bcl9 and analogues was observed.
You can reference link of the kit as following
https://dldevelop.com/Research-reagent/dl-bcl9-mu.html
https://www.dldevelop.com/uploadfile/data/DL-Bcl9-Mu.pdf
Introduction
| Item | Standard | Test | |
| Description |
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of Bcl9 in mouse 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 Bcl9 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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