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Human Apoptosis Inducing Factor (AIF) ELISA Kit

Human Apoptosis Inducing Factor (AIF) ELISA Kit  AIF DL-AIF-Hu PDCD8 Programmed Cell Death protein 8 Apoptosis-inducing factor 1 mitochondrial
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Human Apoptosis Inducing Factor (AIF) ELISA Kit  AIF DL-AIF-Hu PDCD8 Programmed Cell Death protein 8 Apoptosis-inducing factor 1 mitochondrial 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 Apoptosis Inducing Factor (AIF) ELISA Kit
Method: Sandwich
Synonyms:

PDCD8; Programmed Cell Death protein 8; Apoptosis-inducing factor 1, mitochondrial

Detection range: 0.156-10ng/mL
Target Protein: AIF
Size: 96T/48T
Quality guarantee period: for 12 months, 16 months
Catalog number: DL-AIF-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-AIF-Hu.pdf DL-AIF-Hu.pdf
Human Apoptosis Inducing Factor (AIF) ELISA Kit elisa kit elisa kits
1.Overview:

Other names:PDCD8; Programmed Cell Death protein 8; Apoptosis-inducing factor 1, mitochondrial

Function: Probable oxidoreductase that has a dual role in controlling cellular life and death; during apoptosis, it is translocated from the mitochondria to the nucleus to function as a proapoptotic factor in a caspase-independent pathway, while in normal mitochondria, it functions as an antiapoptotic factor via its oxidoreductase activity. The soluble form (AIFsol) found in the nucleus induces 'parthanatos' i.e., caspase-independent fragmentation of chromosomal DNA. Interacts with EIF3G,and thereby inhibits the EIF3 machinery and protein synthesis, and activates casapse-7 to amplify apoptosis. Plays a critical role in caspase-independent, pyknotic cell death in hydrogen peroxide-exposed cells. Binds to DNA in a sequence-independent manner.

Sequence:

 10         20         30         40         50
 MFRCGGLAAG ALKQKLVPLV RTVCVRSPRQ RNRLPGNLFQ RWHVPLELQM
         60         70         80         90        100
 TRQMASSGAS GGKIDNSVLV LIVGLSTVGA GAYAYKTMKE DEKRYNERIS
        110        120        130        140        150
 GLGLTPEQKQ KKAALSASEG EEVPQDKAPS HVPFLLIGGG TAAFAAARSI
        160        170        180        190        200
 RARDPGARVL IVSEDPELPY MRPPLSKELW FSDDPNVTKT LRFKQWNGKE
        210        220        230        240        250
 RSIYFQPPSF YVSAQDLPHI ENGGVAVLTG KKVVQLDVRD NMVKLNDGSQ
        260        270        280        290        300
 ITYEKCLIAT GGTPRSLSAI DRAGAEVKSR TTLFRKIGDF RSLEKISREV
        310        320        330        340        350
 KSITIIGGGF LGSELACALG RKARALGTEV IQLFPEKGNM GKILPEYLSN
        360        370        380        390        400
 WTMEKVRREG VKVMPNAIVQ SVGVSSGKLL IKLKDGRKVE TDHIVAAVGL
        410        420        430        440        450
 EPNVELAKTG GLEIDSDFGG FRVNAELQAR SNIWVAGDAA CFYDIKLGRR
        460        470        480        490        500
 RVEHHDHAVV SGRLAGENMT GAAKPYWHQS MFWSDLGPDV GYEAIGLVDS
        510        520        530        540        550
 SLPTVGVFAK ATAQDNPKSA TEQSGTGIRS ESETESEASE ITIPPSTPAV
        560        570        580        590        600
 PQAPVQGEDY GKGVIFYLRD KVVVGIVLWN IFNRMPIARK IIKDGEQHED
        610
 LNEVAKLFNI HED   
2.Features
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of AIF in human serum, plasma, tissue homogenates 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 AIF is typically less than 0.064ng/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 AIF.
No significant cross-reactivity or interference between AIF and analogues was observed.
 
You can reference link of the kit as following
https://dldevelop.com/Research-reagent/dl-aif-hu.html
https://dldevelop.com/Research-reagent/dl-aif-ra.html
 
 
 
Introduction
ItemStandardTest
Description

The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of AIF in human serum, plasma, tissue homogenates and 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 AIF 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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