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Human ATPase, Ca++ Transporting, Plasma Membrane 2 (ATP2B2) ELISA Kit

Human ATPase, Ca++ Transporting, Plasma Membrane 2 (ATP2B2) ELISA Kit DL-ATP2B2-Hu ATP2B2 PMCA2 Plasma Membrane Ca2+ Pump 2 Plasma Membrane Calcium-Transporting ATPase 2
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Human ATPase, Ca++ Transporting, Plasma Membrane 2 (ATP2B2) ELISA Kit DL-ATP2B2-Hu ATP2B2 PMCA2 Plasma Membrane Ca2+ Pump 2 Plasma Membrane Calcium-Transporting ATPase 2 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 ATPase, Ca++ Transporting, Plasma Membrane 2 (ATP2B2) ELISA Kit
Method: Sandwich
Synonyms:

PMCA2; Plasma Membrane Ca2+ Pump 2; Plasma Membrane Calcium-Transporting ATPase 2

Detection range: 0.312-20ng/mL
Target Protein: ATP2B2
Size: 96T/48T
Quality guarantee period: for 12 months, 16 months
Catalog number: DL-ATP2B2-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-ATP2B2-Hu.pdf DL-ATP2B2-Hu.pdf
Human ATPase, Ca++ Transporting, Plasma Membrane 2 (ATP2B2) ELISA Kit elisa kit elisa kits
1.Overview:

Other names:PMCA2; Plasma Membrane Ca2+ Pump 2; Plasma Membrane Calcium-Transporting ATPase 2

Function: This magnesium-dependent enzyme catalyzes the hydrolysis of ATP coupled with the transport of calcium out of the cell.

Sequence:                                                                                                                                                                  

        10         20         30         40         50
MGDMTNSDFY SKNQRNESSH GGEFGCTMEE LRSLMELRGT EAVVKIKETY
        60         70         80         90        100
GDTEAICRRL KTSPVEGLPG TAPDLEKRKQ IFGQNFIPPK KPKTFLQLVW
       110        120        130        140        150
EALQDVTLII LEIAAIISLG LSFYHPPGEG NEGCATAQGG AEDEGEAEAG
       160        170        180        190        200
WIEGAAILLS VICVVLVTAF NDWSKEKQFR GLQSRIEQEQ KFTVVRAGQV
       210        220        230        240        250
VQIPVAEIVV GDIAQVKYGD LLPADGLFIQ GNDLKIDESS LTGESDQVRK
       260        270        280        290        300
SVDKDPMLLS GTHVMEGSGR MLVTAVGVNS QTGIIFTLLG AGGEEEEKKD
       310        320        330        340        350
KKGVKKGDGL QLPAADGAAA SNAADSANAS LVNGKMQDGN VDASQSKAKQ
       360        370        380        390        400
QDGAAAMEMQ PLKSAEGGDA DDRKKASMHK KEKSVLQGKL TKLAVQIGKA
       410        420        430        440        450
GLVMSAITVI ILVLYFTVDT FVVNKKPWLP ECTPVYVQYF VKFFIIGVTV
       460        470        480        490        500
LVVAVPEGLP LAVTISLAYS VKKMMKDNNL VRHLDACETM GNATAICSDK
       510        520        530        540        550
TGTLTTNRMT VVQAYVGDVH YKEIPDPSSI NTKTMELLIN AIAINSAYTT
       560        570        580        590        600
KILPPEKEGA LPRQVGNKTE CGLLGFVLDL KQDYEPVRSQ MPEEKLYKVY
       610        620        630        640        650
TFNSVRKSMS TVIKLPDESF RMYSKGASEI VLKKCCKILN GAGEPRVFRP
       660        670        680        690        700
RDRDEMVKKV IEPMACDGLR TICVAYRDFP SSPEPDWDNE NDILNELTCI
       710        720        730        740        750
CVVGIEDPVR PEVPEAIRKC QRAGITVRMV TGDNINTARA IAIKCGIIHP
       760        770        780        790        800
GEDFLCLEGK EFNRRIRNEK GEIEQERIDK IWPKLRVLAR SSPTDKHTLV
       810        820        830        840        850
KGIIDSTHTE QRQVVAVTGD GTNDGPALKK ADVGFAMGIA GTDVAKEASD
       860        870        880        890        900
IILTDDNFSS IVKAVMWGRN VYDSISKFLQ FQLTVNVVAV IVAFTGACIT
       910        920        930        940        950
QDSPLKAVQM LWVNLIMDTF ASLALATEPP TETLLLRKPY GRNKPLISRT
       960        970        980        990       1000
MMKNILGHAV YQLALIFTLL FVGEKMFQID SGRNAPLHSP PSEHYTIIFN
      1010       1020       1030       1040       1050
TFVMMQLFNE INARKIHGER NVFDGIFRNP IFCTIVLGTF AIQIVIVQFG
      1060       1070       1080       1090       1100
GKPFSCSPLQ LDQWMWCIFI GLGELVWGQV IATIPTSRLK FLKEAGRLTQ
      1110       1120       1130       1140       1150
KEEIPEEELN EDVEEIDHAE RELRRGQILW FRGLNRIQTQ IRVVKAFRSS
      1160       1170       1180       1190       1200
LYEGLEKPES RTSIHNFMAH PEFRIEDSQP HIPLIDDTDL EEDAALKQNS
      1210       1220       1230       1240
SPPSSLNKNN SAIDSGINLT TDTSKSATSS SPGSPIHSLE TSL     

                               
2.Features
INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of ATP2B2 in human tissue homogenates, cell lysates or 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 ATP2B2 is typically less than 0.112ng/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 ATP2B2.
No significant cross-reactivity or interference between ATP2B2 and analogues was observed.
 
You can reference link of the kit as following
https://dldevelop.com/Research-reagent/dl-atp2b2-hu.html
https://dldevelop.com/Research-reagent/dl-atp2b2-mu.html
https://dldevelop.com/Research-reagent/dl-atp2b2-ra.html
Introduction
ItemStandardTest
Description

The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of ATP2B2 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 ATP2B2 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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