ELISA Kit for Preferentially Expressed Antigen In Melanoma (PRAME)
Sizes: 24T, 48T, 96T
Catalogue Numbers: SEH323Hu-24, SEH323Hu-48, SEH323Hu-96
Citations, Manuals and MSDS Available upon request.
Featured Series: Sandwich kit
Target: Preferentially Expressed Antigen In Melanoma
Alternative Names: MAPE; OIP4; CT130; Cancer/Testis Antigen 130; Opa-interacting protein 4; Melanoma antigen preferentially expressed in tumors
Target Species: Human
Uniprot: P78395
Gene ID: 23532
Featured Series Function: Detects protein (regular version)
Specificity: Reactive with Human PRAME / Preferentially Expressed Antigen In Melanoma
Method: Colormetric
Detection principle: Double-antibody Sandwich
Detection
range: 0.312-20ng/mL
Sensitivity: 0.113ng/mL
Assay Time: 3h
Sample Size: 100uL
Recommended/Predicted
Sample Types: Tissue Homogenates, Cell Lysates, Cell Culture Supernates and other Biological Fluids
Assay Precision: Intra-Assay: CV<10%, Inter-Assay: CV<12%
Storage: 4°C for 1 month/ -20°C for long-term(One year within shelf life)
Shelf-life: 12 months
Specificity: This assay has high sensitivity and excellent specificity for detection of Preferentially Expressed Antigen In Melanoma (PRAME).
No significant cross-reactivity or interference between Preferentially Expressed Antigen In Melanoma (PRAME) and analogues was observed.
Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Preferentially Expressed Antigen In Melanoma (PRAME) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Preferentially Expressed Antigen In Melanoma (PRAME) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
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.
Target: 1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.
Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Preferentially Expressed Antigen In Melanoma (PRAME). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Preferentially Expressed Antigen In Melanoma (PRAME). 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 Preferentially Expressed Antigen In Melanoma (PRAME), 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 Preferentially Expressed Antigen In Melanoma (PRAME) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Research Area: Tumor immunity;
Research Use Only