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Human Amyloid Beta Precursor Like Protein 2 (APLP2) ELISA Kit

Human Amyloid Beta Precursor Like Protein 2 (APLP2) ELISA Kit APLP2 DL-APLP2-Hu APPH APPL2 CDEBP Amyloid protein homolog CDEI box-binding protein
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Human Amyloid Beta Precursor Like Protein 2 (APLP2) ELISA Kit APLP2 DL-APLP2-Hu APPH APPL2 CDEBP Amyloid protein homolog CDEI box-binding protein 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 Amyloid Beta Precursor Like Protein 2 (APLP2) ELISA Kit
Method: Sandwich
Synonyms:

APPH; APPL2; CDEBP; Amyloid protein homolog; CDEI box-binding protein

Detection range: 0.156-10ng/mL
Target Protein: APLP2
Size: 96T/48T
Quality guarantee period: for 12 months, 16 months
Catalog number: DL-APLP2-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-APLP2-Hu.pdf DL-APLP2-Hu.pdf
Human Amyloid Beta Precursor Like Protein 2 (APLP2) ELISA Kit elisa kit elisa kits

1.Overview

Other names:APPH; APPL2; CDEBP; Amyloid protein homolog; CDEI box-binding protein

Function:  May play a role in the regulation of hemostasis. The soluble form may have inhibitory properties towards coagulation factors. May interact with cellular G-protein signaling pathways. May bind to the DNA 5'-GTCACATG-3'(CDEI box). Inhibits trypsin, chymotrypsin, plasmin, factor XIA and plasma and glandular kallikrein.

Sequence:

  50: MAATGTAAAA ATGRLLLLLL VGLTAPALAL AGYIEALAAN AGTGFAVAEP 
 100: QIAMFCGKLN MHVNIQTGKW EPDPTGTKSC FETKEEVLQY CQEMYPELQI     
 150: TNVMEANQRV SIDNWCRRDK KQCKSRFVTP FKCLVGEFVS DVLLVPEKCQ   
 200: FFHKERMEVC ENHQHWHTVV KEACLTQGMT LYSYGMLLPC GVDQFHGTEY     
 250: VCCPQTKIIG SVSKEEEEED EEEEEEEDEE EDYDVYKSEF PTEADLEDFT   
 300: EAAVDEDDED EEEGEEVVED RDYYYDTFKG DDYNEENPTE PGSDGTMSDK      
 350: EITHDVKAVC SQEAMTGPCR AVMPRWYFDL SKGKCVRFIY GGCGGNRNNF      
 400: ESEDYCMAVC KAMIPPTPLP TNDVDVYFET SADDNEHARF QKAKEQLEIR  
 450: HRNRMDRVKK EWEEAELQAK NLPKAERQTL IQHFQAMVKA LEKEAASEKQ  
 500: QLVETHLARV EAMLNDRRRM ALENYLAALQ SDPPRPHRIL QALRRYVRAE  
 550: NKDRLHTIRH YQHVLAVDPE KAAQMKSQVM THLHVIEERR NQSLSLLYKV   
 600: PYVAQEIQEE IDELLQEQRA DMDQFTASIS ETPVDVRVSS EESEEIPPFH  
 650: PFHPFPALPE NEDTQPELYH PMKKGSGVGE QDGGLIGAEE KVINSKNKVD  
 700: ENMVIDETLD VKEMIFNAER VGGLEEERES VGPLREDFSL SSSALIGLLV  
 750: IAVAIATVIV ISLVMLRKRQ YGTISHGIVE VDPMLTPEER HLNKMQNHGY    
 760: ENPTYKYLEQ MQI                                                                   
2.Features
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of APLP2 in human tissue homogenates, cell lysates or other biological fluids.

DETECTION RANGE
0.156-10ng/mL. The standard curve concentrations used for the ELISA’s were 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.312ng/mL, 0.156ng/mL.
 
SENSITIVITY
The minimum detectable dose of APLP2 is typically less than 0.054ng/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 APLP2.
No significant cross-reactivity or interference between APLP2 and analogues was observed.
 
You can reference link of the kit as following
https://dldevelop.com/Research-reagent/dl-aplp2-hu.html
https://www.dldevelop.com/uploadfile/data/DL-APLP2-Hu.pdf 
Introduction
ItemStandardTest
Description

The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of APLP2 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

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 APLP2 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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