- Product Name
- Human AGE(Advanced glycation end-product) ELISA Kit
- Alias
- AGE ELISA Kit, Advanced glycation end-product ELISA Kit
- Catalogue No.
- EH0622
- Size
- 48T/96T
- Species
- Human
- UniProt No.
- P51606
- 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 AGE , no obvious cross reaction with other analogues.
- ELISA Kit Components
Kit Components | Item | Size(48T) | Size(96T) | Storage Condition for Opened Kit |
E001 | ELISA Microplate(Dismountable) | 8×6 | 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 month at -20°C |
E002 | Lyophilized Standard | 1vial | 2vial | Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C |
E003 | Biotin-labeled Antibody(Concentrated, 100X) | 60ul | 120ul | 2-8°C (Avoid Direct Light) |
E034 | HRP-Streptavidin Conjugate(SABC, 100X) | 60ul | 120ul |
E024 | TMB Substrate | 5ml | 10ml |
E039 | Sample Dilution Buffer | 10ml | 20ml | 2-8°C |
E040 | Antibody Dilution Buffer | 5ml | 10ml |
E049 | SABC Dilution Buffer | 5ml | 10ml |
E026 | Stop Solution | 5ml | 10ml |
E038 | Wash Buffer(Concentrated, 25X) | 15ml | 30ml |
E006 | Plate Sealer | 3 pieces | 5 pieces | |
E007 | Product Description | 1 copy | 1 copy |
- Required Instruments and Reagents
-
- Microplate reader (wavelength: 450nm)
- 37°C incubator (CO2 incubator for cell culture is not recommenced.)
- 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
- Assay Procedure Summary
-
- 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.
- Standard Curve
-
This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.)
Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.
STD.(ng/ml) | OD-1 | OD-2 | Average |
0 | 0.058 | 0.061 | 0.059 |
0.312 | 0.196 | 0.206 | 0.2 |
0.625 | 0.288 | 0.302 | 0.294 |
1.25 | 0.469 | 0.492 | 0.478 |
2.5 | 0.815 | 0.857 | 0.832 |
5 | 1.251 | 1.315 | 1.276 |
10 | 1.756 | 1.846 | 1.792 |
20 | 2.134 | 2.243 | 2.178 |
- Recovery
-
Add a certain amount of AGE into the sample. Calculate the recovery by comparing the measured value with the expected amount of AGE in the sample.
Sample Type | Recovery Range(%) | Average(%) |
serum(n=10) | 88-97 | 94 |
EDTA plasma(n=10) | 89-98 | 95 |
Heparin plasma(n=10) | 88-105 | 98 |
- Linearity
-
Dilute the sample with a certain amount of AGE at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type | 1:2 | 1:4 | 1:8 |
serum(n=10) | 86-100% | 82-100% | 80-100% |
EDTA plasma(n=10) | 85-100% | 82-94% | 81-99% |
Heparin plasma(n=10) | 85-102% | 89-93% | 81-96% |
- Precision(%)
-
Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.
Item | Intra-assay Precision | Inter-assay Precision |
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 20 | 20 | 20 |
Mean (ng/ml) | 0.63 | 2.41 | 10.26 | 0.6 | 2.47 | 9.96 |
Standard deviation | 0.03 | 0.14 | 0.55 | 0.03 | 0.12 | 0.5 |
CV(%) | 5.12 | 5.89 | 5.34 | 5.23 | 4.88 | 4.98 |
- 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 |