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Human ATP Binding Cassette Transporter A3 (ABCA3) ELISA Kit

Human ATP Binding Cassette Transporter A3 (ABCA3) ELISA Kit ABCA3 DL-ABCA3-Hu ABC-A3 ABC-C ABC3 ABC-C transporter
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Human ATP Binding Cassette Transporter A3 (ABCA3) ELISA Kit ABCA3 DL-ABCA3-Hu ABC-A3 ABC-C ABC3 ABC-C transporter 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 A3 (ABCA3) ELISA Kit
Method: Sandwich
Synonyms:

ABC-A3; ABC-C; ABC3; ABC-C transporter

Detection range: 78.125-5000pg/mL
Target Protein: ABCA3
Size: 96T/48T
Quality guarantee period: for 12 months, 16 months
Catalog number: DL-ABCA3-Hu (traditional) (ready-to-use)
Assay length 1-4.5Hours 1-3.5Hours
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 -20℃
  • Faster reaction compare to other brands
  • 16 months shelf life
Instruction Manual DL-ABCA3-Hu.pdf DL-ABCA3-Hu.pdf
Human ATP Binding Cassette Transporter A3 (ABCA3) ELISA Kit elisa kit elisa kits
1. Overview

Other names:ABC-A3; ABC-C; ABC3; ABC-C transporter

Function: Plays an important role in the formation of pulmonary surfactant, probably by transporting lipids such as cholesterol.

Sequence
MAVLRQLALL  LWKNYTLQKR  KVLVTVLELF  LPLLFSGILI  WLRLKIQSEN  
VPNATIYPGQ  SIQELPLFFT  FPPPGDTWEL  AYIPSHSDAA  KTVTETVRRA  
LVINMRVRGF  PSEKDFEDYI  RYDNCSSSVL  AAVVFEHPFN  HSKEPLPLAV  
KYHLRFSYTR  RNYMWTQTGS  FFLKETEGWH  TTSLFPLFPN  PGPREPTSPD  
GGEPGYIREG  FLAVQHAVDR  AIMEYHADAA  TRQLFQRLTV  TIKRFPYPPF  
IADPFLVAIQ  YQLPLLLLLS  FTYTALTIAR  AVVQEKERRL  KEYMRMMGLS  
SWLHWSAWFL  LFFLFLLIAA  SFMTLLFCVK  VKPNVAVLSR  SDPSLVLAFL  
LCFAISTISF  SFMVSTFFSK  ANMAAAFGGF  LYFFTYIPYF  FVAPRYNWMT  
LSQKLCSCLL  SNVAMAMGAQ  LIGKFEAKGM  GIQWRDLLSP  VNVDDDFCFG  
QVLGMLLLDS  VLYGLVTWYM  EAVFPGQFGV  PQPWYFFIMP  SYWCGKPRAV  
AGKEEEDSDP  EKALRNEYFE  AEPEDLVAGI  KIKHLSKVFR  VGNKDRAAVR  
DLNLNLYEGQ  ITVLLGHNGA  GKTTTLSMLT  GLFPPTSGRA  YISGYEISQD  
MVQIRKSLGL  CPQHDILFDN  LTVAEHLYFY  AQLKGLSRQK  CPEEVKQMLH  
IIGLEDKWNS  RSRFLSGGMR  RKLSIGIALI  AGSKVLILDE  PTSGMDAISR  
RAIWDLLQRQ  KSDRTIVLTT  HFMDEADLLG  DRIAIMAKGE  LQCCGSSLFL  
KQKYGAGYHM  TLVKEPHCNP  EDISQLVHHH  VPNATLESSA  GAELSFILPR  
ESTHRFEGLF  AKLEKKQKEL  GIASFGASIT  TMEEVFLRVG  KLVDSSMDIQ  
AIQLPALQYQ  HERRASDWAV  DSNLCGAMDP  SDGIGALIEE  ERTAVKLNTG  
LALHCQQFWA  MFLKKAAYSW  REWKMVAAQV  LVPLTCVTLA  LLAINYSSEL 
FDDPMLRLTL  GEYGRTVVPF  SVPGTSQLGQ  QLSEHLKDAL  QAEGQEPREV  
LGDLEEFLIF  RASVEGGGFN  ERCLVAASFR  DVGERTVVNA  LFNNQAYHSP  
ATALAVVDNL  LFKLLCGPHA  SIVVSNFPQP  RSALQAAKDQ  FNEGRKGFDI  
ALNLLFAMAF  LASTFSILAV  SERAVQAKHV  QFVSGVHVAS  FWLSALLWDL  
ISFLIPSLLL  LVVFKAFDVR  AFTRDGHMAD  TLLLLLLYGW  AIIPLMYLMN  
FFFLGAATAY  TRLTIFNILS  GIATFLMVTI  MRIPAVKLEE  LSKTLDHVFL  
VLPNHCLGMA  VSSFYENYET  RRYCTSSEVA  AHYCKKYNIQ  YQENFYAWSA  
PGVGRFVASM  AASGCAYLIL  LFLIETNLLQ  RLRGILCALR  RRRTLTELYT  
RMPVLPEDQD  VADERTRILA  PSPDSLLHTP  LIIKELSKVY  EQRVPLLAVD 
RLSLAVQKGE  CFGLLGFNGA  GKTTTFKMLT  GEESLTSGDA  FVGGHRISSD  
VGKVRQRIGY  CPQFDALLDH  MTGREMLVMY  ARLRGIPERH  IGACVENTLR 
GLLLEPHANK  LVRTYSGGNK  RKLSTGIALI  GEPAVIFLDE  PSTGMDPVAR  
RLLWDTVARA  RESGKAIIIT  SHSMEECEAL  CTRLAIMVQG  QFKCLGSPQH  
LKSKFGSGYS  LRAKVQSEGQ  QEALEEFKAF  VDLTFPGSVL  EDEHQGMVHY  
HLPGRDLSWA  KVFGILEKAK  EKYGVDDYSV  SQISLEQVFL  SFAHLQPPTA  
EEGR

2. Features
 
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of ABCA3 in human 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 ABCA3 is typically less than 29pg/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 ABCA3.
No significant cross-reactivity or interference between ABCA3 and analogues was observed.
Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between ABCA3 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
https://www.dldevelop.com/uploadfile/data/DL-ABCA3-Hu.pdf
Introduction
ItemStandardTest
Description

The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of ABCA3 in human tissue homogenates 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

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 ABCA3 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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