适用生物 oryctolagus cuniculus (rabbit，兔)
检测范围 12.35-1000ng/ml 灵敏度 5.25ng/ml
样本类型 plasma.
实验时长 2.5h 实验方法 竞争抑制法 规格 96t
血管性血友病因子(vwf)检测试剂盒
elisa kit for von willebrand factor (vwf)
for in vitro and research use only, not for use in clinical diagnostic procedures!
organism species oryctolagus cuniculus (rabbit)
product no. cea833rb
sample type plasma.
format 96-well strip plate
assay length 2.5 hours
detection range 12.35-1000ng/ml the standard curve concentrations used for the elisa’s were 1000ng/ml, 333.33ng/ml, 111.11ng/ml, 37.04ng/ml, 12.35ng/ml
sensitivity the minimum detectable dose of this kit is typically less than 5.25ng/ml.
血管性血友病因子(vwf)检测试剂盒specificity
this assay has high sensitivity and excellent specificity for detection of von willebrand factor (vwf).
no significant cross-reactivity or interference between von willebrand factor (vwf) and analogues was observed.
recovery
matrices listed below were spiked with certain level of recombinant von willebrand factor (vwf) and the recovery rates were calculated by comparing the measured value to the expected amount of von willebrand factor (vwf) in samples.
血管性血友病因子(vwf)检测试剂盒
matrix recovery range (%) average(%)
serum(n=5) 84-105 97
edta plasma(n=5) 91-105 94
heparin plasma(n=5) 81-104 89
precision
intra-assay precision (precision within an assay): 3 samples with low, middle and high level von willebrand factor (vwf) were tested 20 times on one plate, respectively.
inter-assay precision (precision between assays): 3 samples with low, middle and high level von willebrand factor (vwf) were tested on 3 different plates, 8 replicates in each plate.
cv(%) = sd/meanx100
intra-assay: cv<10%
inter-assay: cv<12%
linearity
the linearity of the kit was assayed by testing samples spiked with appropriate concentration of von willebrand factor (vwf) and their serial dilutions. the results were demonstrated by the percentage of calculated concentration to the expected.
血管性血友病因子(vwf)检测试剂盒
sample 1:2 1:4 1:8 1:16
serum(n=5) 89-103% 88-97% 80-88% 82-105%
edta plasma(n=5) 92-105% 81-98% 92-99% 98-105%
heparin plasma(n=5) 79-99% 80-101% 90-97% 90-104%
stability
the stability of 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 condition.
to minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. it is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
reagents and materials provided
reagents quantity reagents quantity
pre-coated, ready to use 96-well strip plate 1 plate sealer for 96 wells 4
standard 2 standard diluent 1×20ml
detection reagent a 1 assay diluent a 1×12ml
detection reagent b 1×120μl assay diluent b 1×12ml
reagent diluent 1×300μl stop solution 1×6ml
tmb substrate 1×9ml instruction manual 1
wash buffer (30 × concentrate) 1×20ml
血管性血友病因子(vwf)检测试剂盒assay procedure summary
1. prepare all reagents, samples and standards;
2. add 50μl standard or sample to each well.
and then add 50μl prepared detection reagent a immediay.
shake and mix. incubate 1 hour at 37oc;
3. aspirate and wash 3 times;
4. add 100μl prepared detection reagent b. incubate 30 minutes at 37oc;
5. aspirate and wash 5 times;
6. add 90μl substrate solution. incubate 15-25 minutes at 37oc;
7. add 50μl stop solution. read at 450 nm immediay.
test principle
this assay employs the competitive inhibition enzyme immunoassay technique. a monoclonal antibody specific to von willebrand factor (vwf) has been pre-coated onto a microplate. a competitive inhibition reaction is launched between biotin labeled von willebrand factor (vwf) and unlabeled von willebrand factor (vwf) (standards or samples) with the pre-coated antibody specific to von willebrand factor (vwf). after incubation the unbound conjugate is washed off. next, avidin conjugated to horseradish peroxidase (hrp) is added to each microplate well and incubated. the amount of bound hrp conjugate is reverse proportional to the concentration of von willebrand factor (vwf) in the sample. after addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of von willebrand factor (vwf) in the sample.