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Human ATP Binding Cassette Transporter C11 (ABCC11) ELISA Kit

Human ATP Binding Cassette Transporter C11 (ABCC11) ELISA Kit ABC-C11 DL-ABCC11-Hu ABC-C11 MRP8 EWWD Multidrug resistance-associated protein 8
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Human ATP Binding Cassette Transporter C11 (ABCC11) ELISA Kit ABC-C11 DL-ABCC11-Hu ABC-C11 MRP8 EWWD Multidrug resistance-associated protein 8 code image

two product lines: Traditional ELISA Kit and Ready-to-Use ELISA Kit.

Traditional ELISA Kit Ready-to-Use ELISA KIT
Product name: Human ATP Binding Cassette Transporter C11 (ABCC11) ELISA Kit
Method: Competition
Synonyms:

ABC-C11; MRP8; EWWD; Multidrug resistance-associated protein 8

Detection range: 0.312-20ng/mL
Reactivity: ABCC11
Size: 96T/48T
Quality guarantee period: for 12 months
Catalog number: DL-ABCC11-Hu (traditional) (ready-to-use)
Assay length 1-2.5Hours 1-2.5Hours
Advantages:
  • Competitive price.
  • High sensitivity.
  • High stability.
  • 12 months shelf life.
  • Pre-diluted Detection Reagent A and B
  • Reduction in the number of steps when conducting the test
  • All the reagents can be stored at 4℃
  • Faster reaction compare to other brands
  • 12 months shelf life
Instruction Manual DL-ABCC11-Hu.pdf DL-ABCC11-Hu.pdf
Human ATP Binding Cassette Transporter C11 (ABCC11) ELISA Kit elisa kit elisa kits
1. Overview

Other names:ABC-C11; MRP8; EWWD; Multidrug resistance-associated protein 8

Function: Participates in physiological processes involving bile acids, conjugated steroids and cyclic nucleotides. Enhances the cellular extrusion of cAMP and cGMP. Stimulates the ATP-dependent uptake of a range of physiological and synthetic lipophilic anions, including the glutathione S-conjugates leukotriene C4 and dinitrophenyl S-glutathione, steroid sulfates such as dehydroepiandrosterone 3-sulfate (DHEAS) and estrone 3-sulfate, glucuronides such as estradiol 17-beta-D-glucuronide (E(2)17betaG), the monoanionic bile acids glycocholate and taurocholate, and methotrexate. Probably functions to secrete earwax.

Sequence
MTRKRTYWVP  NSSGGLVNRG  IDIGDDMVSG  LIYKTYTLQD  GPWSQQERNP  
EAPGRAAVPP  WGKYDAALRT  MIPFRPKPRF  PAPQPLDNAG  LFSYLTVSWL  
TPLMIQSLRS  RLDENTIPPL  SVHDASDKNV  QRLHRLWEEE  VSRRGIEKAS  
VLLVMLRFQR  TRLIFDALLG  ICFCIASVLG  PILIIPKILE  YSEEQLGNVV  
HGVGLCFALF  LSECVKSLSF  SSSWIINQRT  AIRFRAAVSS  FAFEKLIQFK  
SVIHITSGEA  ISFFTGDVNY  LFEGVCYGPL  VLITCASLVI  CSISSYFIIG  
YTAFIAILCY  LLVFPLAVFM  TRMAVKAQHH  TSEVSDQRIR  VTSEVLTCIK  
LIKMYTWEKP  FAKIIEDLRR  KERKLLEKCG  LVQSLTSITL  FIIPTVATAV  
WVLIHTSLKL  KLTASMAFSM  LASLNLLRLS  VFFVPIAVKG  LTNSKSAVMR  
FKKFFLQESP  VFYVQTLQDP  SKALVFEEAT  LSWQQTCPGI  VNGALELERN  
GHASEGMTRP  RDALGPEEEG  NSLGPELHKI  NLVVSKGMML  GVCGNTGSGK  
SSLLSAILEE  MHLLEGSVGV  QGSLAYVPQQ  AWIVSGNIRE  NILMGGAYDK  
ARYLQVLHCC  SLNRDLELLP  FGDMTEIGER  GLNLSGGQKQ  RISLARAVYS  
DRQIYLLDDP  LSAVDAHVGK  HIFEECIKKT  LRGKTVVLVT  HQLQYLEFCG  
QIILLENGKI  CENGTHSELM  QKKGKYAQLI  QKMHKEATSD  MLQDTAKIAE  
KPKVESQALA  TSLEESLNGN  AVPEHQLTQE  EEMEEGSLSW  RVYHHYIQAA  
GGYMVSCIIF  FFVVLIVFLT  IFSFWWLSYW  LEQGSGTNSS  RESNGTMADL  
GNIADNPQLS  FYQLVYGLNA  LLLICVGVCS  SGIFTKVTRK  ASTALHNKLF  
NKVFRCPMSF  FDTIPIGRLL  NCFAGDLEQL  DQLLPIFSEQ  FLVLSLMVIA  
VLLIVSVLSP  YILLMGAIIM  VICFIYYMMF  KKAIGVFKRL  ENYSRSPLFS 
HILNSLQGLS  SIHVYGKTED  FISQFKRLTD  AQNNYLLLFL  SSTRWMALRL  
EIMTNLVTLA  VALFVAFGIS  STPYSFKVMA  VNIVLQLASS  FQATARIGLE  
TEAQFTAVER  ILQYMKMCVS  EAPLHMEGTS  CPQGWPQHGE  IIFQDYHMKY  
RDNTPTVLHG  INLTIRGHEV  VGIVGRTGSG  KSSLGMALFR  LVEPMAGRIL  
IDGVDICSIG  LEDLRSKLSV  IPQDPVLLSG  TIRFNLDPFD  RHTDQQIWDA  
LERTFLTKAI  SKFPKKLHTD  VVENGGNFSV  GERQLLCIAR  AVLRNSKIIL  
IDEATASIDM  ETDTLIQRTI  REAFQGCTVL  VIAHRVTTVL  NCDHILVMGN  
GKVVEFDRPE  VLRKKPGSLF  AALMATATSS  LR
 
2.  Features
 
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of ABCC11 in human tissue homogenates, cell lysates and 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 ABCC11 is typically less than 0.103ng/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 ABCC11.
No significant cross-reactivity or interference between ABCC11 and analogues was observed.
Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between ABCC11 and all analogues, therefore, cross reactivity may still exist.

IMPORTANT NOTES
1. Limited by the current conditions and scientific technology, it is impossible to conduct comprehensive identification and analysis tests on the raw materials provided by suppliers. As a result, it is possible there are some qualitative and/or technical risks.
2. The final experimental results will be closely related to the validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available to obtain accurate results.
3. Kits from different batches may be a little different in detection range, sensitivity and color developing time. Please perform the experiment exactly according to the instruction manual included in your kit. Electronic ones on our website are for reference only.
4. Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
5. Protect all reagents from strong light during storage and incubation. All bottle caps of reagents should be closed tightly to prevent evaporation of liquids and contamination by microorganisms.
6. There may be a foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
7. Incorrect procedures during reagent preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
8. Even the same experimenter may get different results from two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before the general assay for each batch is recommended.
9. Each kit has undergone several rigorous quality control tests. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipment. Intra-assay variance among kits from different batches could arise from the above factors as well.
10. Kits from different manufacturers with the same item might produce different results, since we have not compared our products with other manufacturers.
11. The standard of the kit and immunogen used for antibody preparation are commonly recombinant proteins. Different expressed sequence, expression systems, purification methods might be used in recombinant protein preparation. Besides, there might exist differences on the screening technique of antibody and antibody pairs in our kit. Thus we can not guarantee the kit could detect recombinant protein from other companies. So, it is not recommended to use the kit for the detection of recombinant protein.
12. Validity period: 12 months.
13. The instruction manual also works with the 48T kit, but all reagents in the 48T kit are reduced by half.
 
PRECAUTION
The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this reagent.

You can reference link of the kit as following
http://www.dldevelop.com/uploadfile/data/DL-ABCC11-Hu.pdf
Introduction
Item Standard Test
Description

The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of ABCC11 in human tissue homogenates, cell lysates or other biological fluids.

Conform
IdentificationColorimetricPositive
Composition Traditional ELISA Kit Ready-to-Use ELISA KITConform
Pre-coated, ready to use 96-well strip plate 1Pre-coated, ready to use 96-well strip plate 1
Plate sealer for 96 wells 2Plate sealer for 96 wells 2
Standard 2 Standard 2
Diluents buffer 1×45mLStandard Diluent 1×20mL
Detection Reagent A 1×120μLDetection Solution A 1×12mL
Detection Reagent B 1×120μLDetection Solution B 1×12mL
TMB Substrate 1×9mLTMB Substrate 1×9mL
Stop Solution 1×6mLStop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mLWash Buffer (30 × concentrate) 1×20mL
Instruction manual 1 Instruction manual 1

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to the index has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled the index and unlabeled the index (Standards or samples) with the pre-coated antibody specific to the index. After incubation the unbound conjugate is washed off. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of the index in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of the index in the sample.

Recovery

Matrices listed below were spiked with certain level of recombinant the index 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 condition.
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 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37℃;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 1 hour at 37℃;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37℃;
7. Add 50µL Stop Solution. Read at 450 nm immediately.
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