Human TNF Receptor Associated Factor 3 (TRAF3) ELISA Kit

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

	Traditional ELISA Kit	Ready-to-Use ELISA KIT
Product name:	Human TNF Receptor Associated Factor 3 (TRAF3) ELISA Kit
Method:	Sandwich
Synonyms:	LAP1; CAP-1; CD40bp; CRAF1; CD40 receptor-associated factor 1; CD40-binding protein; LMP1-associated protein 1
Detection range:	78.125-5,000pg/mL
Target Protein:	TRAF3
Size:	96T/48T
Quality guarantee period:	for 12 months, 16 months
Catalog number:	DL-TRAF3-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

Human TNF Receptor Associated Factor 3 (TRAF3) ELISA Kit

1.Overview：

Other names：LAP1; CAP-1; CD40bp; CRAF1; CD40 receptor-associated factor 1; CD40-binding protein; LMP1-associated protein 1

Function: Regulates pathways leading to the activation of NF-kappa-B and MAP kinases, and plays a central role in the regulation of B-cell survival. Part of signaling pathways leading to the production of cytokines and interferon. Required for normal antibody isotype switching from IgM to IgG. Plays a role T-cell dependent immune responses. Plays a role in the regulation of antiviral responses. Is an essential constituent of several E3 ubiquitin-protein ligase complexes. May have E3 ubiquitin-protein ligase activity and promote 'Lys-63'-linked ubiquitination of target proteins. Inhibits activation of NF-kappa-B in response to LTBR stimulation. Inhibits TRAF2-mediated activation of NF-kappa-B. Down-regulates proteolytic processing of NFKB2, and thereby inhibits non-canonical activation of NF-kappa-B. Promotes ubiquitination and proteasomal degradation of MAP3K14.

Sequence：

        10         20         30         40         50
 MESSKKMDSP GALQTNPPLK LHTDRSAGTP VFVPEQGGYK EKFVKTVEDK
         60         70         80         90        100
 YKCEKCHLVL CSPKQTECGH RFCESCMAAL LSSSSPKCTA CQESIVKDKV
        110        120        130        140        150
 FKDNCCKREI LALQIYCRNE SRGCAEQLML GHLLVHLKND CHFEELPCVR
        160        170        180        190        200
 PDCKEKVLRK DLRDHVEKAC KYREATCSHC KSQVPMIALQ KHEDTDCPCV
        210        220        230        240        250
 VVSCPHKCSV QTLLRSELSA HLSECVNAPS TCSFKRYGCV FQGTNQQIKA
        260        270        280        290        300
 HEASSAVQHV NLLKEWSNSL EKKVSLLQNE SVEKNKSIQS LHNQICSFEI
        310        320        330        340        350
 EIERQKEMLR NNESKILHLQ RVIDSQAEKL KELDKEIRPF RQNWEEADSM
        360        370        380        390        400
 KSSVESLQNR VTELESVDKS AGQVARNTGL LESQLSRHDQ MLSVHDIRLA
        410        420        430        440        450
 DMDLRFQVLE TASYNGVLIW KIRDYKRRKQ EAVMGKTLSL YSQPFYTGYF
        460        470        480        490        500
 GYKMCARVYL NGDGMGKGTH LSLFFVIMRG EYDALLPWPF KQKVTLMLMD
        510        520        530        540        550
 QGSSRRHLGD AFKPDPNSSS FKKPTGEMNI ASGCPVFVAQ TVLENGTYIK
        560
 DDTIFIKVIV DTSDLPDP

2．Features
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of TRAF3 in human serum, plasma, tissue homogenates, cell lysates or other biological fluids.

DETECTION RANGE
78.1-5000pg/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 TRAF3 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 TRAF3.
No significant cross-reactivity or interference between TRAF3 and analogues was observed.

You can reference link of the kit as following
https://dldevelop.com/Research-reagent/dl-traf3-hu.html
https://www.dldevelop.com/uploadfile/data/DL-TRAF3-Hu.pdf

Introduction

Item	Standard		Test
Description	The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of TRAF3 in human serum, plasma, tissue homogenates, cell lysates 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 TRAF3 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.