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Human free-IgE (free-Immunoglobulin E) ELISA Kit (AEFI01097)

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SKU:
AEFI01097
Product Type:
ELISA Kit
Size:
96 Assays
Sensitivity:
0.188ng/ml
Range:
0.313-20ng/ml
$719

Description

Product Name: Human free-IgE (free-Immunoglobulin E) ELISA Kit
Alias: free-IgE ELISA Kit, free-Immunoglobulin E ELISA Kit
SKU: AEFI01097
Size: 96T
Species: Human
Sample Type: Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
Detection Method: Sandwich ELISA, Double Antibody
Detection Wavelength: OD450
Reaction Duration: 4 hours
Range: 0.313-20ng/ml
Sensitivity: 0.188ng/ml
Storage: 2-8°C(Sealed), Don't cryopreserve.
Specificity: Specifically binds with free-IgE , no obvious cross reaction with other analogues.
Component Size (96T) Storage
ELISA Microplate (Dismountable) 8×12 Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 months at -20°C
Lyophilized Standard 2 vials Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 months at -20°C
Biotin-labeled Antibody (Concentrated, 100X) 120 ul 2-8°C (Avoid Direct Light)
HRP-Streptavidin Conjugate (SABC, 100X) 120 ul
TMB Substrate 10 ml
Sample Dilution Buffer 20 ml 2-8°C
Antibody Dilution Buffer 10 ml
SABC Dilution Buffer 10 ml
Stop Solution 10 ml
Wash Buffer (Concentrated, 25X) 30 ml
Plate Sealer 5 pieces
Product Description 1 copy

Other materials and equipment required:

  • Microplate reader (wavelength: 450nm)
  • 37°C incubator (CO2 incubator for cell culture is not recommended.)
  • Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
  • Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
  • Sterile tubes and Eppendorf tubes with disposable tips
  • Absorbent paper and loading slot
  • Deionized or distilled water
Recovery: Add a certain amount of free-IgE into the sample. Calculate the recovery by comparing the measured value with the expected amount of free-IgE in the sample.
Matrix Recovery range(%) Average(%)
serum(n=5) 85-102 94
EDTA plasma(n=5) 88-102 95
Heparin plasma(n=5) 85-97 93
Linearity: Dilute the sample with a certain amount of free-IgE at 1:2, 1:4 and 1:8 to get the recovery range.
Sample 1:2 1:4 1:8
serum(n=5) 86-103% 83-101% 84-95%
EDTA plasma(n=5) 93-104% 82-100% 88-100%
Heparin plasma(n=5) 89-105% 88-98% 80-91%
Stability: Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.
ELISA kit(n=5) 37°C for 1 month 2-8°C for 6 months
Average(%) 80 95-100
Precision(%): Intra-Assay: CV<8%
Inter-Assay: CV<10%
Step 1: Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.
Washing: Wash the plate twice without immersing.
Step 2: Add 100ul biotin-antibody working solution, seal the plate and statically incubate for 60 minutes at 37°C.
Washing: Wash the plate three times and immerse for 1min each time.
Step 3: Add 100ul HRP-Streptavidin Conjugate (SABC) working solution, seal the plate and statically incubate for 30 minutes at 37°C.
Washing: Wash the plate five times and immerse for 1min each time.
Step 4: Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
Step 5: Add 50ul stop solution. Read at 450nm immediately and calculate.

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